TWI608848B - Liposome formulations - Google Patents

Description

脂質體調配物 Liposomal formulation 相關申請案之交叉參考Cross-reference to related applications

本申請案主張對分別於2012年8月21日；2013年3月15日；及2013年8月5日申請之美國臨時專利申請案61/691,455、61/791,693及61/862,300之優先權，該等申請案全部以引用的方式全文併入本文中。 This application claims priority to US Provisional Patent Applications Nos. 61/691,455, 61/791,693 and 61/862,300, filed on August 21, 2012; March 15, 2013; and August 5, 2013, respectively. All of these applications are hereby incorporated by reference in their entirety.

本發明係關於包含通常藉由玻璃體內注射施加至眼睛之眼用醫藥及容許將該玻璃體內醫藥局部施加至眼睛之脂質體遞送劑的醫藥調配物。本發明亦係關於抗血管生成化合物，例如選自(例如)蘭尼單抗(ranibizumab)之單株抗體或其片段，蘭尼單抗係抑制VEGF之作用之血管內皮生長因子結合劑；及/或多重激酶VEGFR及PDGFR抑制劑，例如舒尼替尼(sunitinib)；及選自醫藥上可接受之脂質體之遞送劑。該等調配物係用於治療動物及人類之各種血管生成病症及疾病，且較佳用於選自年齡相關黃斑退化、糖尿病黃斑水腫及角膜新血管生成之眼病。 The present invention relates to pharmaceutical formulations comprising ophthalmic drugs that are typically applied to the eye by intravitreal injection and liposome delivery agents that permit topical application of the intravitreal drug to the eye. The present invention also relates to an anti-angiogenic compound, for example, a monoclonal antibody selected from, for example, ranibizumab or a fragment thereof, and ranibizumab is a vascular endothelial growth factor binding agent that inhibits the action of VEGF; Or a multi-kinase VEGFR and PDGFR inhibitor, such as sunitinib; and a delivery agent selected from pharmaceutically acceptable liposomes. Such formulations are useful in the treatment of various angiogenic disorders and diseases in animals and humans, and are preferably used in ophthalmopathy selected from the group consisting of age-related macular degeneration, diabetic macular edema, and corneal neovascularization.

眼用疾病治療通常需要將特定藥物局部投與或玻璃體內注射至眼睛，此取決於特定疾病或病狀及關於特定藥物及疾病之投與途徑之效力。在某些眼睛疾病或病狀中，僅在該藥物係藉由玻璃體內注射投與時，可達成有效治療。藉由玻璃體內注射可有效治療多種眼睛疾病 及病狀。同時，該等注射可引起包括眼睛感染(眼內炎)、眼睛發炎、視網膜脫落及眼壓增加在內之嚴重副效應及/或與其相關。由於該等副效應或風險，眼睛之局部治療已成為治療眼睛病狀之藥物之較佳投與途徑及聖杯(Holy Grail)，此乃因在幾乎所有情形下，藥物之局部投與不能有效治療某些眼睛病狀、尤其發生在眼睛後部之彼等病狀。因此，相關技藝需要研發有效治療該等病狀並消除進行玻璃體內注射之需要之調配物。本發明者相信其已發現此一媒劑。美國專利第6,884,879號揭示各種抗VEGF抗體。此專利具體闡述並主張經批准並以商標名稱LUCENTIS®出售之單株抗體蘭尼單抗。其中所揭示之抗體係闡述為能夠預防、逆轉及/或緩和各種疾病之症狀，並闡述為具有抑制內皮細胞之VEGF誘導之增生之能力及抑制VEGF誘導之血管生成之能力。LUCENTIS®已經批准以0.5mg(0.05mL)之劑量強度藉由一個月一次玻璃體內注射用於新生血管(濕性)年齡相關黃斑退化(AMD)，且儘管有效性下降，但可以在至少四個月之一個月一次之初始方案後每三個月注射一次來投與所述經批准治療。LUCENTIS®亦在美國經批准按一個月一次玻璃體內方案使用0.5mg(0.05mL)用於視網膜靜脈阻塞(RVO)之後之黃斑水腫。另外，LUCENTIS®在歐洲及美國經批准以可注射調配物形式用於糖尿病黃斑水腫。 The treatment of ophthalmic diseases usually requires topical or intravitreal injection of a particular drug to the eye, depending on the particular disease or condition and the efficacy of the particular drug and the route of administration of the disease. In certain ocular diseases or conditions, effective treatment can be achieved only when the drug is administered by intravitreal injection. Effective intravitreal injection for a variety of eye diseases And the condition. At the same time, such injections may cause and/or be associated with severe side effects including eye infections (endophthalmitis), eye irritation, retinal detachment, and increased intraocular pressure. Due to these side effects or risks, topical treatment of the eye has become the preferred route of administration for the treatment of ocular conditions and the Holy Grail, because in almost all cases, local administration of the drug cannot be effectively treated. Certain eye conditions, especially those that occur in the back of the eye. Accordingly, the related art requires the development of formulations that are effective in treating such conditions and eliminating the need for intravitreal injection. The inventors believe that this agent has been found. Various anti-VEGF antibodies are disclosed in U.S. Patent No. 6,884,879. This patent specifically states and claims a monoclonal antibody, ranibizumab, approved and sold under the trade name LUCENTIS®. The anti-system disclosed therein is described as capable of preventing, reversing, and/or alleviating the symptoms of various diseases, and is described as having the ability to inhibit VEGF-induced proliferation of endothelial cells and inhibit VEGF-induced angiogenesis. LUCENTIS® has been approved for intravitreal injection once a month for neovascular (wet) age-related macular degeneration (AMD) at a dose intensity of 0.5 mg (0.05 mL), and may be at least four in spite of reduced efficacy The approved treatment is administered every three months after the initial protocol once a month. LUCENTIS® is also approved in the United States for a one-month intravitreal regimen using 0.5 mg (0.05 mL) for macular edema after retinal vein occlusion (RVO). In addition, LUCENTIS® is approved for use in the preparation of injectable formulations for diabetic macular edema in Europe and the United States.

舒尼替尼(SU-11248，舒癌特(Sutent))係在2006年1月由FDA批准為用於治療轉移性腎細胞癌及胃腸基質瘤之單一療法。舒尼替尼抑制至少8種受體蛋白-酪胺酸激酶，該等受體蛋白-酪胺酸激酶包括血管內皮生長因子受體1至3(VEGFR1至VEGFR3)、血小板源生長因子受體(PDGFRα及PDGFRβ)。此化合物藉由減少藉助VEGFR1、VEGFR2及PDGFRβ之信號傳導來抑制血管生成。PDGFRβ係發現於圍繞毛細管內皮細胞之外周細胞中¹(Roskoski，2007，附帶PDF文檔)。有證據證明，抗PDGF之組合使用在一些VEGF相關之疾病之治療中優於蘭尼 單抗單一療法。 Sunitinib (SU-11248, Sutent) was approved by the FDA in January 2006 as a monotherapy for the treatment of metastatic renal cell carcinoma and gastrointestinal stromal tumors. Sunitinib inhibits at least eight receptor proteins, tyrosine kinases, including vascular endothelial growth factor receptors 1 to 3 (VEGFR1 to VEGFR3), platelet-derived growth factor receptors ( PDGFRα and PDGFRβ). This compound inhibits angiogenesis by reducing signaling by VEGFR1, VEGFR2 and PDGFRβ. PDGFRβ Department found around the outside of capillary endothelial cells in peripheral cells ¹ (Roskoski, 2007, comes with a PDF document). There is evidence that the combination of anti-PDGF is superior to ranibizumab monotherapy in the treatment of some VEGF-related diseases.

存在多種副效應或潛在副效應，包括與玻璃體內注射抗VEGF抗體及其他眼用藥物相關之患者不適。玻璃體內注射程序需要具有一般無菌規則之專用無塵室；復蘇設施必須立即可用。此程序之併發症包括感染性眼內炎；視網膜脫落及創傷性白內障。玻璃體內注射之其他可能併發症包括眼內壓力變化，尤指眼內壓力升高。可能在注射任一種類之醫藥後立即發生與注射相關之眼內壓力升高，及可能在注射後數天或甚至數月後檢測到明確之與藥物相關之眼內壓力變化。參見Semin.Ophthamol.2009 Mar-Apr；24(2)：100-5。 There are a variety of side effects or potential side effects, including patient discomfort associated with intravitreal injection of anti-VEGF antibodies and other ophthalmic drugs. Intravitreal injection procedures require a dedicated clean room with general aseptic rules; the resuscitation facility must be available immediately. Complications of this procedure include infectious endophthalmitis; retinal detachment and traumatic cataracts. Other possible complications of intravitreal injection include changes in intraocular pressure, especially intraocular pressure. An intraocular pressure increase associated with injection may occur immediately after injection of any type of medication, and a clear drug-related intraocular pressure change may be detected several days or even months after the injection. See Semin. Ophthamol. 2009 Mar-Apr; 24(2): 100-5.

相關技藝對該等抗VEGF抗體及諸如以下等其他有效眼用藥物之新局部治療方案存在迫切且未得到滿足之醫學需求：抗微生物劑、抗病毒劑、皮質類固醇及抗血管內皮生長因子藥劑，其係藉助玻璃體內注射投與之主要藥物類別。本發明包含該等脂質體與該等藥物類別內之任一該等藥物之呈局部調配物形式之組合。儘管一般而言美國專利第6,884,879號揭示包括脂質體之各種可能遞送方法或試劑及包括局部投與該等VEGF單株抗體之各種投與途徑，但蘭尼單抗之唯一經批准形式係存於液體調配物中之玻璃體內形式。相關技藝需要有效治療患有包括眼病之VEGF相關之病症之人的局部蘭尼單抗調配物。 Related Art There is an urgent and unmet medical need for such anti-VEGF antibodies and new topical treatments for other effective ophthalmic drugs such as: antimicrobial agents, antiviral agents, corticosteroids, and anti-vascular endothelial growth factor agents, It is the main drug class that is administered by intravitreal injection. The invention encompasses combinations of such liposomes with a topical formulation of any of the classes of such drugs. Although generally, U.S. Patent No. 6,884,879 discloses various possible delivery methods or reagents including liposomes and various routes of administration including local administration of antibodies to such VEGF monoclonal antibodies, the only approved form of ranibizumab is present in Intravitreal form in liquid formulations. The related art requires a topical ranibizumab formulation that is effective in treating a subject having a VEGF-related disorder including an eye disease.

本發明者已滿足此未得到滿足之需求，且已驚奇地發現某些包含該等VEGF單株抗體之脂質體調配物，且某些脂質體有效緩解患有糖尿病黃斑水腫之患者。可向受影響眼睛有效地局部投與該等調配物，且其較局部施加玻璃體內調配物有效。美國專利第6,958,160號揭示並主張自成型之熱力學穩定之脂質體。美國專利公開案第2010/0076209號(以引用的方式併入本文中)揭示用於脂質體及藥物遞送之PEG-脂質偶聯物。包括該等以商標名稱Qsomes^TM(在下文中，Qsome)出售之自成型之熱力學穩定之脂質體的各種基於脂質之產物 係由Biozone Laboratories出售用於多種治療用途並經由各種投與途徑(包括藉由向皮膚局部投與)。本發明者已發現，可將包含蘭尼單抗及該等自成型之熱力學穩定之脂質體及/或PEG-脂質偶聯物之醫藥組合物局部遞送至需要治療VEGF相關之眼用疾病及病狀之患者之眼睛。本發明進一步包含包含本文中所陳述之Qsome(自成型之熱力學穩定之脂質體)及藥物之局部調配物，其中該調配物適宜於治療眼後段疾病及/或為組合眼前段/後段疾病之疾病。所主張調配物適宜於局部投與，並特別用於治療通常經由眼周途徑或經由玻璃體內投與(眼內遞送)治療之眼用疾病及病狀。眼周投與包括結膜下、特農囊下、眼球後及球周投與。存在一些已揭示為能夠藉由局部投與遞送至眼後段之藥物-該等包括ESBA105，即抗TNF-α單鏈抗體；***(dexamethasone)；奈帕芬胺(nepafenac)；美金剛HCl(memantine HCl)；多佐胺(dorzolamide)；溴莫尼定(brimonidine)及倍他洛爾(betaxolol)。據信，該等藥物以Qsome調配物形式之局部投與將導致更有效局部遞送及更有效眼後段遞送。較佳地，然而，所主張之發明包含包括本文中所闡述之Qsome脂質體及迄今為止仍未有效地藉助局部投與遞送之藥物的局部調配物。 The inventors have met this unmet need and have surprisingly discovered certain liposome formulations comprising such VEGF monoclonal antibodies, and certain liposomes are effective in alleviating patients with diabetic macular edema. The formulations can be effectively administered topically to the affected eye and are more effective than topical application of the intravitreal formulation. U.S. Patent No. 6,958,160 discloses and claims a thermodynamically stable liposome from self-forming. U.S. Patent Publication No. 2010/0076209, which is incorporated herein by reference, discloses PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT PCT Patent No They include those under the trade name Qsomes ^TM (hereinafter, Qsome) molded from a variety of thermodynamically stable of-sale liposomes of lipid-based system products sold by Biozone Laboratories for various therapeutic purposes and via various routes of administration (including by Partially administered to the skin). The present inventors have discovered that a pharmaceutical composition comprising ranibizumab and such self-forming thermodynamically stable liposomes and/or PEG-lipid conjugates can be delivered topically to ophthalmic diseases and diseases in need of treatment for VEGF The eyes of the patient. The invention further comprises a topical formulation comprising a Qsome (self-forming thermodynamically stable liposome) and a medicament as set forth herein, wherein the formulation is suitable for treating a disease of the posterior segment of the eye and/or a disease associated with a combination of anterior/posterior segment disease . The claimed formulations are suitable for topical administration and are particularly useful for the treatment of ophthalmic diseases and conditions which are typically administered via the periocular route or via intravitreal administration (intraocular delivery). Periocular injections include subconjunctival, special undergrowth, posterior eyeball, and peribulbar. There are some drugs that have been disclosed to be deliverable to the posterior segment of the eye by topical administration - these include ESBA 105, an anti-TNF-α single chain antibody; dexamethasone; nepafenac; memantine HCl (memantine HCl); dozolamide; brimonidine and betaxolol. It is believed that topical administration of such drugs in the form of a Qsome formulation will result in more effective topical delivery and more effective delivery of the posterior segment of the eye. Preferably, however, the claimed invention comprises a topical formulation comprising a Qsome liposome as set forth herein and a drug that has heretofore not been effectively administered by topical delivery.

通常藉由玻璃體內注射給予之藥物包括抗微生物劑、抗病毒劑、皮質類固醇及抗血管內皮生長因子藥劑。本發明包含脂質體調配物，該脂質體調配物包含自成型之熱力學穩定之脂質體及選自抗微生物劑、抗病毒劑、皮質類固醇及抗血管內皮生長因子藥劑中之任一者或其組合之活性醫藥劑，其中該調配物適宜於局部遞送至患者之眼睛以治療眼部疾病或病狀。待克菲那(Diclofenac)、加替沙星(gatifloxacin)、乳酸司帕沙星(sparfloxacin lactate)、GCV、地美環素(demeclocycline)、夫比普洛芬(flurbiprofen)、多柔比星(doxorubicin)、塞來昔布(celecoxib)、布***(budesonide)及順鉑 (cisplatin)已調配成膠質劑型用於經角膜或經鞏膜遞送。相對於陰離子型及中性脂質體，含有青黴素G、托平卡胺(tropicamide)及乙醯唑胺(acetazolamide)之陽離子型脂質體已用於提供跨越角膜之最大藥物運輸。參見Schaeffer等人，Liposomes in topical drug deliver.Invest.Ophthalmol.Vis Sci.1982：22(2)：220-7；Nagarsenker等人，Preparation and evaluation of liposomal formulations of tropicamide for ocular delivery.Int J Pharma.1999：190(1)：63-71及Hathout等人，Liposome as an ocular delivery system for acetazolamide：in vitro and in vivo studies.AAPS PharmaSciTech.2007；8(1)：1。 The drugs usually administered by intravitreal injection include an antimicrobial agent, an antiviral agent, a corticosteroid, and an anti-vascular endothelial growth factor agent. The present invention comprises a liposome formulation comprising a self-forming thermodynamically stable liposome and any one or combination of the group consisting of an antimicrobial agent, an antiviral agent, a corticosteroid, and an anti-vascular endothelial growth factor agent. An active pharmaceutical agent, wherein the formulation is suitable for topical delivery to the eye of a patient to treat an ocular disease or condition. Wait for Diclofenac, gatifloxacin, sparfloxacin lactate, GCV, demeclocycline, flurbiprofen, doxorubicin ( Doxorubicin), celecoxib, budesonide and cisplatin (cisplatin) has been formulated into a glial dosage form for delivery via the cornea or transscleral. Cationic liposomes containing penicillin G, tropicamide and acetazolamide have been used to provide maximum drug transport across the cornea relative to anionic and neutral liposomes. See Schaeffer et al, Liposomes in topical drug deliver. Invest. Ophthalmol. Vis Sci. 1982: 22(2): 220-7; Nagarsenker et al, Preparation and evaluation of liposomal formulations of tropicamide for ocular delivery. Int J Pharma. 1999 : 190(1): 63-71 and Hathout et al, Liposome as an ocular delivery system for acetazolamide: in vitro and in vivo studies. AAPS PharmaSciTech. 2007; 8(1): 1.

糖尿病視網膜病變(DR)係糖尿病之最常見大血管併發症²(Fong,2004)，且係工作年齡成年人中視覺喪失新個案之主要原因。糖尿病黃斑水腫(DME)係患有DR之患者之視覺喪失之最常見原因。DME之盛行率係3%最新診斷，其中每年約75,000個DME新個案(USA)。全世界患有糖尿病之患者數係令人驚愕之285,000,000。DME係由一系列生物化學及細胞變化造成，該等變化最終引起漸進洩漏及滲出，從而導致視網膜增厚及黃斑中央之1個視神經盤直徑內之硬滲出物。DME係患有糖尿病之患者之受損視覺之一最常見原因³(Bhagat,2009)。在2年之隨訪後，大約50%之患者經歷2行之BCVA之喪失⁴(Meyer,2007)。 Diabetic retinopathy (DR) is the most common major vascular complication of diabetes ² (Fong, 2004) and is the leading cause of new cases of visual loss in working-age adults. Diabetic macular edema (DME) is the most common cause of visual loss in patients with DR. The prevalence of DME is 3% of the latest diagnosis, with approximately 75,000 new DME cases (USA) per year. The number of patients with diabetes worldwide is 285,000,000. DME is caused by a series of biochemical and cellular changes that eventually cause progressive leakage and exudation, resulting in thickening of the retina and hard exudate within the diameter of one optic disc in the center of the macula. DME is one of the most common causes of impaired vision in patients with diabetes ³ (Bhagat, 2009). After 2 years of follow-up, approximately 50% of patients experienced The loss of BCVA 2 lines of ⁴ (Meyer, 2007).

在DME中，受損血管將流體洩漏至導致腫脹之視網膜(黃斑)之中央部分中。黃斑與敏銳之中央視覺有涉及。中央窩係在黃斑之中央。DME可發生在患有1型或2型糖尿病之患者中。美國大約26,000,000個人患有糖尿病，且每年20歲或更大之人中診斷出1,900,000個新個案。估計每年發生多達75.000個DME新案例。DME係大部分發達國家中工作年齡之群體之失明之主要原因。DME之第一線療法係雷射外科手術，該雷射外科手術密封漏的血管以減小流體洩漏並減少視網膜中之 流體量。因此，相關技藝顯著需要產生向該等患者提供治療緩解之新調配物。 In DME, damaged blood vessels leak fluid into the central portion of the retina (the macula) that causes swelling. The macula is involved with the keen central vision. The central fossa is in the center of the macula. DME can occur in patients with type 1 or type 2 diabetes. Approximately 26 million people in the United States have diabetes, and 1,900,000 new cases are diagnosed among people aged 20 or older each year. It is estimated that as many as 75.000 DME new cases occur each year. DME is the leading cause of blindness in working age groups in most developed countries. The first line of therapy for DME is laser surgery, which seals leaking blood vessels to reduce fluid leakage and reduce retinal The amount of fluid. Accordingly, there is a significant need in the art to create new formulations that provide therapeutic relief to such patients.

本發明係包含抗血管生成化合物或其他眼用活性成份及脂質體之醫藥調配物。較佳抗血管生成化合物係抗VEGF抗體。較佳地以局部調配物形式向需要治療之患者之眼睛投與該醫藥調配物。該醫藥調配物係用於治療VEGF相關之眼病，包括(例如)年齡相關黃斑退化及糖尿病黃斑水腫。本發明亦係關於適宜於局部投與且有效治療角膜新生血管疾病之調配物。 The invention is a pharmaceutical formulation comprising an anti-angiogenic compound or other ophthalmic active ingredient and liposomes. Preferred anti-angiogenic compounds are anti-VEGF antibodies. The pharmaceutical formulation is preferably administered to the eye of the patient in need of treatment in the form of a topical formulation. The pharmaceutical formulation is for the treatment of VEGF-related eye diseases including, for example, age-related macular degeneration and diabetic macular edema. The invention is also directed to formulations suitable for topical administration and effective treatment of corneal neovascular diseases.

較佳抗VEGF抗體係選自蘭尼單抗(LUCENTIS®)，且可如美國專利第6,884,879號中所闡述製備，該專利以引用的方式併入本文中。此產物係以處方玻璃體內液體調配物之形式出售。蘭尼單抗係重組人類化IgG1 κ同型單株抗體片段。此抗體結合至VEGF-A並抑制其，且具有大約48千道耳頓(kilodalton)之分子量。該產物係於含有四環素之營養培養基中之大腸桿菌(E.coli)表現系統中產生。該處方產物係於含有0.05mL之10mg/mL蘭尼單抗之單次使用玻璃小瓶中供應。諸如貝伐珠單抗(bevacizumab)等其他抗VEGF抗體亦可用於治療某些疾病及病狀，例如角膜新生血管疾病。本文中所揭示之局部脂質體調配物有助於整個抗體(例如貝伐珠單抗)滲透至具有異常新脈管系統之眼睛中。 A preferred anti-VEGF anti-system is selected from the group consisting of ranibizumab (LUCENTIS®) and can be prepared as set forth in U.S. Patent No. 6,884,879, incorporated herein by reference. This product is sold as a prescription intravitreal liquid formulation. Ranibizumab is a recombinant humanized IgG1 κ homologous antibody fragment. This antibody binds to and inhibits VEGF-A and has a molecular weight of approximately 48 kilodaltons. This product was produced in an E. coli expression system in a nutrient medium containing tetracycline. The formulation product was supplied in a single use glass vial containing 0.05 mL of 10 mg/mL ranibizumab. Other anti-VEGF antibodies, such as bevacizumab, can also be used to treat certain diseases and conditions, such as corneal neovascular diseases. The topical liposome formulations disclosed herein facilitate penetration of whole antibodies (e.g., bevacizumab) into the eye with an abnormally new vasculature.

用於本發明之脂質體較佳包含美國專利第6,958,160號中所闡述之彼等脂質體，該專利以引用的方式全文併入本文中。如其中所闡述，脂質體係自封閉膠質顆粒，其中由一或多個脂質雙層構成之膜囊封一部分其中懸浮該等脂質體之水溶液。如'160專利中亦陳述，並非所有脂質體皆相同，且實際上，脂質體可具有包括以下之問題：膠質不穩定性及因極端條件(例如升高之壓力及溫度以及高剪切條件-其全 部皆可降解脂質組份)導致之製造問題。與脂質體相關之其他問題通常可包括雙層之大小及數量之異質分佈，其可引起或加劇放大問題。另外，滅菌條件亦可產生關於脂質體之問題。脂質體亦可具有因在懸浮液中時聚集而導致之膠質不穩定性。此導致融合問題，且對於此問題，解決方案通常係凍乾，其係昂貴之額外步驟。'160專利中所揭示之脂質體已克服該等問題，且驚奇地發現該等特定脂質體在與諸如蘭尼單抗等抗VEGF抗體組合時係有效的。 Liposomes for use in the present invention preferably comprise the liposomes as set forth in U.S. Patent No. 6,958,160, the disclosure of which is incorporated herein in its entirety. As set forth therein, the lipid system self-blocks colloidal particles wherein a membrane comprised of one or more lipid bilayers encapsulates a portion of an aqueous solution in which the liposomes are suspended. As also stated in the '160 patent, not all liposomes are identical, and in fact, liposomes can have problems including gel instability and extreme conditions (e.g., elevated pressure and temperature, and high shear conditions - Its full The manufacturing process can be caused by the degradation of the lipid component. Other problems associated with liposomes can generally include heterogeneous distributions of the size and number of bilayers that can cause or exacerbate amplification problems. In addition, sterilization conditions can also cause problems with liposomes. Liposomes can also have colloidal instability due to aggregation during suspension. This leads to a fusion problem, and for this problem, the solution is usually lyophilized, which is an extra step of being expensive. The liposomes disclosed in the '160 patent have overcome these problems and have surprisingly found that such specific liposomes are effective when combined with an anti-VEGF antibody such as ranibizumab.

特定而言，包含自成型之熱力學穩定之脂質體及抗VEGF抗體之調配物特別適宜於在VEGF相關之眼用疾病及病狀之治療中局部施加。 In particular, formulations comprising self-forming thermodynamically stable liposomes and anti-VEGF antibodies are particularly suitable for topical application in the treatment of VEGF-related ophthalmic diseases and conditions.

用於本發明調配物之脂質體包含二醯基甘油-PEG化合物。該等化合物之熔點係低於約40℃，且醯基鏈之長度為大於或等於約14個碳。如'160專利中所陳述製備該等化合物。較佳脂質PEG偶聯物係PEG-12-GDM(聚乙二醇12-甘油二肉豆蔻酸酯)。PEG藉由在脂質體之外表面產生空間障壁來穩定脂質體，PEG鏈具有介於約300道耳頓與5000道耳頓之間之分子量。脂質體製備需要僅將脂質與水溶液混合。該等種類之脂質體以脂質可存在之最低能態存在，而在水溶液中，脂質體形成之重現性不成問題。 Liposomes for use in the formulations of the invention comprise a dimercaptoglycerol-PEG compound. The compounds have a melting point below about 40 ° C and a thiol chain length of greater than or equal to about 14 carbons. Such compounds are prepared as set forth in the '160 patent. A preferred lipid PEG conjugate is PEG-12-GDM (polyethylene glycol 12-glyceryl dimyristate). PEG stabilizes liposomes by creating a spatial barrier on the outer surface of the liposome, which has a molecular weight between about 300 Daltons and 5000 Daltons. Liposomal preparation requires mixing only the lipid with the aqueous solution. These types of liposomes are present in the lowest energy state in which lipids can be present, whereas in aqueous solutions, the reproducibility of liposome formation is not a problem.

每次當與相同水溶液混合時，經界定之脂質、脂質混合物或脂質/化合物混合物將形成類似脂質體。在臨界濃度(約20重量對體積%)以上，非脂質體結構將開始在水溶液中形成。該等脂質體以其能態最低存在，且係熱力學穩定之自成型脂質體。PEG-12 GDM形成極小囊泡，故其可經滅菌過濾。可向脂質溶液及水溶液提供諸如振動或渦旋等動能。本發明調配物亦可使用6,958,160專利中所通常或具體闡述之其他脂質-PEG偶聯物。另外，亦可使用其他自成型之熱力學穩定之脂質PEG偶聯物，包括美國專利公開案第2010/0076209號中所闡述之 彼等化合物。下表1闡述某些PEG-12 GDM特徵。 A defined lipid, lipid mixture or lipid/compound mixture will form a similar liposome each time when mixed with the same aqueous solution. Above the critical concentration (about 20 wt.% by volume), the non-liposomal structure will begin to form in aqueous solution. These liposomes are minimally present in their energy state and are thermodynamically stable self-forming liposomes. PEG-12 GDM forms very small vesicles, so it can be sterilized and filtered. Kinetic energy such as vibration or vortex can be supplied to the lipid solution and the aqueous solution. Other lipid-PEG conjugates typically or specifically described in the 6,958,160 patent may also be used in the formulations of the invention. In addition, other self-forming thermodynamically stable lipid PEG conjugates can be used, including those described in U.S. Patent Publication No. 2010/0076209. These compounds. Table 1 below illustrates certain PEG-12 GDM characteristics.

分子量：1068g/mol Molecular weight: 1068g/mol

最優pH：5至7 Optimal pH: 5 to 7

溶解性：可溶於有機溶劑中。 Solubility: Soluble in organic solvents.

在本發明之蘭尼單抗局部調配物中，使用基於溶液之最終體積1重量體積%之PEG-12 GDM，且局部溶液之pH為5.5，且脂質體在適當工作範圍中。 In the ranibizumab topical formulation of the invention, PEG-12 GDM based on a final volume of solution of 1 wt%, and the pH of the topical solution was 5.5, and the liposomes were in the appropriate working range.

除抗血管生成化合物(例如，抗VEGF抗體)及熱力學穩定之自成型脂質體以外，該調配物可進一步包含其他醫藥上可接受之賦形劑。該等較佳賦形劑係選自由適宜於向眼睛局部投與之賦形劑組成之群。該等包括表面活性劑、緩衝劑、pH調整劑、鹽及其他該等成份。 In addition to anti-angiogenic compounds (e.g., anti-VEGF antibodies) and thermodynamically stable self-forming liposomes, the formulation may further comprise other pharmaceutically acceptable excipients. These preferred excipients are selected from the group consisting of excipients suitable for topical administration to the eye. These include surfactants, buffers, pH adjusters, salts, and other such ingredients.

該等調配物係用於治療VEGF相關之疾病及病狀及/或其他血管生成病狀。因此，本發明包括該等調配物治療年齡相關黃斑退化、包括糖尿病黃斑水腫之糖尿病視網膜疾病及角膜新血管生成之用途。在較佳實施例中，本發明包含局部調配物，並涵蓋藉由向需要治療之患者之眼睛局部投與該等調配物來治療該等VEGF相關之疾病及病狀之方法。 Such formulations are useful for treating VEGF-related diseases and conditions and/or other angiogenic conditions. Accordingly, the invention includes the use of such formulations to treat age-related macular degeneration, diabetic retinal diseases including diabetic macular edema, and corneal neovascularization. In a preferred embodiment, the invention encompasses topical formulations and encompasses methods of treating such VEGF-related diseases and conditions by topical administration of such formulations to the eye of a patient in need of treatment.

將在以下圖中闡述本發明。 The invention will be illustrated in the following figures.

圖1顯示來自利用脂質體蘭尼單抗調配物治療之單一患者之數 據，該患者在每天給予6滴並持續2週後在8週時期內具有經改良之使用光學同調斷層掃描術量測之中央窩厚度(OCT-CFT)。在第8週，該患者顯示CFT之增加，且在10週時以每天2滴之日劑量再起始治療。圖1顯示CFT在12週至14週時期再次出現改良。 Figure 1 shows the number of single patients from treatment with liposomal ranibizumab formulations According to this, the patient had a modified foveal thickness (OCT-CFT) measured by optical coherence tomography in an 8-week period after giving 6 drops per day for 2 weeks. At week 8, the patient showed an increase in CFT and at 10 weeks the treatment was restarted at a daily dose of 2 drops per day. Figure 1 shows that the CFT reappeared again during the 12 to 14 week period.

圖2顯示利用脂質體蘭尼單抗調配物治療之相同臨床患者，其在每天給予6滴並持續2週後在8週時期內具有經改良之視覺敏銳度量測(ETDRS BCVA)。在第8週時，該患者顯示VA之降低，且在10週時以每天2滴之日劑量再起始治療。圖2顯示VA在12週至14週時期再次出現改良。 Figure 2 shows the same clinical patient treated with the liposomal ranibizumab formulation with a modified visual acuity metric (ETDRS BCVA) over a period of 8 weeks after 6 drops per day for 2 weeks. At week 8, the patient showed a decrease in VA and at 10 weeks the treatment was restarted at a daily dose of 2 drops per day. Figure 2 shows that VA showed improvement again during the 12 to 14 week period.

圖3顯示多層脂質體及單層脂質體之顯微鏡照片。 Figure 3 shows a photomicrograph of multilamellar liposomes and unilamellar liposomes.

圖4顯示利用蘭尼單抗之脂質體調配物治療之患者之中央窩厚度隨時間之OCT結果。 Figure 4 shows the OCT results of foveal thickness over time in patients treated with liposome formulations of ranibizumab.

圖5顯示患者對側眼睛之中央窩厚度隨時間之OCT結果。 Figure 5 shows the OCT results of the foveal thickness over time in the contralateral eye of the patient.

圖6顯示三個患者之所研究眼睛之BCVA隨時間之結果。 Figure 6 shows the results of BCVA over time for the eyes of the three patients studied.

圖7顯示對側眼睛之BCVA隨時間之結果。 Figure 7 shows the results of BCVA over time in the contralateral eye.

本發明係關於醫藥調配物及其用途，其中該較佳調配物包含抗VEGF抗體及自成型之熱力學穩定之脂質體。本發明亦係關於包含抗VEGF抗體及自成型之熱力學穩定之脂質體之局部調配物。本發明進一步包含治療VEGF相關之疾病或病狀之方法，該方法包含向需要治療之患者投與包含抗VEGF抗體及自成型之熱力學穩定之脂質體之調配物。在較佳實施例中，該VEGF相關之疾病或病狀係選自糖尿病視網膜病變(DR)。 The present invention relates to pharmaceutical formulations and uses thereof, wherein the preferred formulation comprises an anti-VEGF antibody and a self-forming thermodynamically stable liposome. The invention is also directed to topical formulations comprising an anti-VEGF antibody and a thermodynamically stable liposome from self-forming. The invention further comprises a method of treating a VEGF-related disease or condition, the method comprising administering to a patient in need of treatment a formulation comprising an anti-VEGF antibody and a self-forming thermodynamically stable liposome. In a preferred embodiment, the VEGF-related disease or condition is selected from the group consisting of diabetic retinopathy (DR).

儘管脂質體「通常」之闡述係與各種活性成份之遞送相關，但相關技藝並未揭示或教示自成型之熱力學穩定之脂質體與抗VEGF抗體之組合。該調配物之脂質體係特別適宜於向需要治療VEGF相關之 疾病或病狀(特定而言眼部疾病或病狀)之患者遞送抗VEGF抗體。本發明之脂質體調配物係特別適於向需要治療(例如)糖尿病黃斑水腫或年齡相關黃斑退化或角膜新血管生成之患者之眼睛局部投與。本發明之脂質體具有使其尤其適於該等局部調配物之期望基本性質。該等脂質體懸浮液在調配物之溫度下熱力學穩定。構成脂質體之脂質之組合物具有若干基本性質。該等脂質具有允許形成脂質體之堆疊參數。該等脂質包括在懸浮液中空間上穩定脂質體之聚乙二醇(PEG)或具有類似性質之聚合物，作為頭基之一部分。另外，該等脂質體具有使其在與水或水溶液混合時呈液體形式之熔融溫度。 Although the "usual" description of liposomes is related to the delivery of various active ingredients, the related art does not disclose or teach the combination of thermodynamically stable liposomes from anti-VEGF antibodies. The lipid system of the formulation is particularly suitable for the treatment of VEGF An anti-VEGF antibody is delivered to a patient with a disease or condition, in particular an ocular disease or condition. The liposome formulations of the present invention are particularly suitable for topical administration to the eye of a patient in need of treatment, for example, diabetic macular edema or age-related macular degeneration or corneal neovascularization. The liposomes of the present invention have desirable basic properties that make them particularly suitable for such topical formulations. The liposomal suspensions are thermodynamically stable at the temperature of the formulation. Compositions of lipids constituting liposomes have several basic properties. These lipids have stacking parameters that allow for the formation of liposomes. These lipids include polyethylene glycol (PEG) or a polymer of similar nature that sterically stabilizes the liposome in suspension as part of the head group. Additionally, the liposomes have a melting temperature that allows them to be in liquid form when mixed with water or an aqueous solution.

如6,610,322專利案中所述，當在水溶液中形成脂質體懸浮液時，需要添加較少能量或不需要添加能量。在本發明中，該較佳方法涉及在含有活性成份-抗VEGF抗體之水溶液之存在下形成脂質體懸浮液。因此，自組裝較佳在具有該活性成份之情況下發生，而非在將活性成份添加至該懸浮液中之前發生。脂質分子分散並自組裝至自然低能態。該等脂質體形成大或小單層囊泡(SUV)或多層囊泡(MLV)(參見圖3)，且如Biozone Laboratories網站中針對Qusomes^TM所闡述。 As described in the 6,610,322 patent, when a liposome suspension is formed in an aqueous solution, less energy needs to be added or no energy needs to be added. In the present invention, the preferred method involves forming a liposome suspension in the presence of an aqueous solution containing the active ingredient-anti-VEGF antibody. Thus, self-assembly preferably occurs with the active ingredient rather than prior to the addition of the active ingredient to the suspension. Lipid molecules disperse and self-assemble to a naturally low energy state. Such liposomes form large or small unilamellar vesicles (SUV) or multilamellar vesicles (MLV) (see FIG. 3), and as set forth Biozone Laboratories site for Qusomes ^TM.

PEG鏈較佳具有介於約300道耳頓與5000道耳頓之間之分子量。適宜脂質之實例包括PEG-12 GDO(甘油二油酸酯)及PEG-12 GDM(甘油二肉豆蔻酸酯)。PEG-12 GDM在25℃下為流體，且具有分別0.853及0.889之堆疊參數P_a及P_v。該等脂質中之每一者在20℃、37℃及60℃下皆形成自發性脂質體。Pa可在介於0.84與0.88之間之範圍內，且Pv介於約0.88與0.93之間。較佳地，適宜化合物形成脂質體代替(例如)微胞。另外，脂質組合物應具有介於約0℃與100℃之間之相轉變溫度-脂質組合物具有允許該組合物在與水溶液混合時呈液體形式之熔融溫度。同樣，該組合物之彎曲彈性模數應使得脂質組合物可在水性環境中形成脂質體，而不需要任何能量輸入或任何顯著能量輸入。可 將動能施加至該溶液。較佳彎曲彈性模數係介於約0kt與15kt之間。彎曲彈性模數在很大程度上係由主鏈決定，且甘油係本發明之較佳主鏈，但亦可使用任一在彎曲彈性模數及適宜官能性方面等效之主鏈。最終溶液中之脂質之相對重量百分比可在大於0wt%(w/w)至約20wt%(w/w)之範圍內。該範圍可介於約1wt%與15wt%之間或介於約1%與10%之間或介於約1% wt/wt與5% wt/wt之間或介於大於0% wt/wt與4% wt/wt之間。 The PEG chain preferably has a molecular weight of between about 300 Daltons and 5,000 Daltons. Examples of suitable lipids include PEG-12 GDO (glycerol dioleate) and PEG-12 GDM (glyceryl dimyristate). PEG-12 GDM is fluid at 25 ° C and has stacking parameters P _a and P _{v of} 0.853 and 0.889, respectively. Each of these lipids forms spontaneous liposomes at 20 ° C, 37 ° C and 60 ° C. Pa can be in the range between 0.84 and 0.88, and Pv is between about 0.88 and 0.93. Preferably, suitable compounds form liposomes in place of, for example, micelles. Additionally, the lipid composition should have a phase transition temperature between about 0 ° C and 100 ° C - the lipid composition has a melting temperature that allows the composition to be in liquid form when mixed with an aqueous solution. Again, the flexural modulus of elasticity of the composition should be such that the lipid composition can form liposomes in an aqueous environment without requiring any energy input or any significant energy input. Kinetic energy can be applied to the solution. Preferably, the flexural modulus of elasticity is between about 0 kt and 15 kt. The flexural modulus of elasticity is largely determined by the backbone, and glycerol is the preferred backbone of the present invention, but any backbone that is equivalent in terms of flexural modulus of elasticity and suitable functionality can be used. The relative weight percentage of lipid in the final solution can range from greater than 0 wt% (w/w) to about 20 wt% (w/w). The range may be between about 1 wt% and 15 wt% or between about 1% and 10% or between about 1% wt/wt and 5% wt/wt or between greater than 0% wt/wt Between 4% wt/wt.

其他分子與具有長於12個之PEG鏈之脂質之混合物亦可用於本發明，限制條件係其形成脂質體。例如，PEG-45 GDS(甘油二硬脂酸酯)與膽固醇之混合物形成脂質體。如'322專利中所闡述，熟習此項技術者可改變包括PEG鏈長度及組成之變量以製備熱力學穩定之自由形成之脂質體，且該等改變在與抗VEGF抗體組合時包括於本發明之範圍內。當在脂質體形成之前添加至脂質中時膽固醇之量係高達約10% w/w。 Mixtures of other molecules with lipids having more than 12 PEG chains can also be used in the present invention, with the proviso that they form liposomes. For example, a mixture of PEG-45 GDS (glycerol distearate) and cholesterol forms a liposome. As described in the '322 patent, those skilled in the art can vary the variables including the length and composition of the PEG chain to prepare thermodynamically stable free-formed liposomes, and such alterations are included in the present invention when combined with an anti-VEGF antibody. Within the scope. The amount of cholesterol is up to about 10% w/w when added to the lipid prior to liposome formation.

除'322專利中所闡述之脂質以外，其他脂質亦可用於本發明。美國專利公開案第2010/0076209號闡述形成適宜於所具體闡述活性成份之藥物遞送之脂質體的某些PEG-脂質偶聯物。該參考文獻中無關於抗VEGF抗體之遞送之教示或參考文獻。特定而言，'209公開案中所闡述之二醯基甘油-聚乙二醇化合物可與抗VEGF抗體組合用於本發明調配物中。脂質化合物之一般結構係顯示於'209公開案中，且包括具有式(R2)(R1)甘油-X-PEG-R1及/或R1-PEG-X-甘油(R2)(R1)之化合物，其中R1較佳係-OH或-OCH3；R2及R3係脂肪酸，包括(但不限於)月桂酸酯、油酸酯、肉豆蔻酸酯、棕櫚酸酯、硬脂酸酯及亞麻油酸酯；且X代表在脂質與PEG之間之單一連接體或複製連接體或兩個或更多個連接體之組合。R2及R3可相同或不同。若R2係在甘油之C1位置處，則R3可定位於C2或C3處。該一般結構包括所有外消旋體及結 構異構體及/或其功能等效物。 In addition to the lipids set forth in the '322 patent, other lipids can also be used in the present invention. U.S. Patent Publication No. 2010/0076209 describes certain PEG-lipid conjugates which form liposomes suitable for the specific delivery of the drug delivery of the active ingredient. There are no teachings or references in this reference regarding the delivery of anti-VEGF antibodies. In particular, the dimethyl glycerol-polyethylene glycol compound set forth in the '209 publication can be used in combination with an anti-VEGF antibody in the formulations of the invention. A general structure of a lipid compound is shown in the '209 publication and includes a compound having the formula (R2) (R1) glycerol-X-PEG-R1 and/or R1-PEG-X-glycerol (R2) (R1), Wherein R1 is preferably -OH or -OCH3; R2 and R3 fatty acids including, but not limited to, lauric acid esters, oleates, myristates, palmitates, stearates, and linolenate; And X represents a single linker or a replication linker or a combination of two or more linkers between the lipid and PEG. R2 and R3 may be the same or different. If R2 is at the C1 position of glycerol, then R3 can be positioned at C2 or C3. The general structure includes all racemates and knots Isomers and/or their functional equivalents.

R1亦可選自(例如)-NH2、-COOH、-OCH2CH3、-OCH2CH2CH3、-OCH2CH2OH、-COCH=CH2、-OCH2CH2NH2、-OSO2CH3、-OCH2C6H6、-OCH2COCH2CH2COONC4H4O2、-CH2CH2=CH2及-OC6H6。R1亦可係有助於將治療劑或靶向劑連接至脂質體表面之官能團。該等官能團可包括胺基烷基酯、馬來醯亞胺、縮水甘油醚、丙酸馬來醯亞胺基、胺基甲酸甲酯、對甲苯磺醯腙鹽、疊氮化物、炔丙基-胺、炔丙基醇、NHS酯、醯肼、琥珀醯亞胺基酯、酒石酸琥珀醯亞胺基酯、琥珀酸琥珀醯亞胺基酯及對甲苯磺酸酯鹽。本發明包括在脂質體組合物內具有抗VEGF抗體之脂質體調配物，且可進一步包括任一其他治療化合物，包括經由R1官能團共價結合或連接至脂質體之抗VEGF抗體。在該例子中，本發明將包括或係組合調配物或具有非共價結合之活性成份及共價結合之活性成份之調配物二者，其中該活性物可相同或不同。 R1 may also be selected from, for example, -NH2, -COOH, -OCH2CH3, -OCH2CH2CH3, -OCH2CH2OH, -COCH=CH2, -OCH2CH2NH2, -OSO2CH3, -OCH2C6H6, -OCH2COCH2CH2COONC4H4O2, -CH2CH2=CH2, and -OC6H6. R1 may also be a functional group that facilitates attachment of a therapeutic or targeting agent to the surface of the liposome. Such functional groups may include aminoalkyl esters, maleimide, glycidyl ether, maleic acid imide, methyl carbazate, p-toluenesulfonate, azide, propargyl - amines, propargyl alcohols, NHS esters, hydrazine, amber succinimide esters, amber succinimide tartrate, amber succinimide succinate and p-toluene sulfonate. The invention includes liposome formulations having an anti-VEGF antibody in a liposome composition, and may further comprise any other therapeutic compound, including an anti-VEGF antibody covalently bound or linked to the liposome via an R1 functional group. In this example, the invention will comprise or be a combination of a formulation or a formulation having a non-covalently bound active ingredient and a covalently bound active ingredient, wherein the actives may be the same or different.

可用於或適宜於此類型之脂質體調配物之連接體包括'209公開案中所闡述之彼等，且該公開案以飲用的方式併入本文中。其中表1闡述或列示該等適宜連接體，且該等連接體包括胺基、琥珀醯基胺基乙醯胺基、2-胺基戊醯胺基、2(2’)-R’-胺基乙醯基等。在每一情形中，脂質在20℃及37℃下形成自發性脂質體，如'209公開案之表4中所顯示。因此，本發明包括闡述為以下之彼等脂質：PEG-12-N1-GDO；PEG-23-N2-GDO；PEG-18-N3-GDO；PEG-23-N4-GDO；PEG-8-S1-GDO；PEG-18-S2-GDO；PEG-12-S3-GDO；PEG-18-Ac1-GDO；PEG-12-Ac2-GDO；PEG-12-N1-GDM；PEG-12-N1-GDLO；PEG-12-S3-GDM；PeG-12-S3-GDLO；PEG-12-Ac2-GDM；PEG-12-Ac2-GDLO；PEG-23-N1-GDL；PEG-12kN1-GDP；PEG-23-Ac2-GDL及PEG-12-Ac2-GDP。GDLO意指甘油二亞麻油酸酯，且GDP意指甘油二棕櫚酸 酯。該等化合物中之每一者在25℃下為流體，且具有在0.830至0.869之範圍內之堆疊參數Pa及在0.872至0.924之範圍內之Pv。 Linkers useful or suitable for liposome formulations of this type include those set forth in the '209 publication, and the disclosure is herein incorporated by reference. Wherein Table 1 illustrates or lists the suitable linkers, and the linkers include an amine group, amber decylaminoethylamino group, 2-aminopentylammonium, 2(2')-R'- Aminoethyl thiol and the like. In each case, the lipid formed spontaneous liposomes at 20 ° C and 37 ° C as shown in Table 4 of the '209 publication. Accordingly, the invention includes the following lipids: PEG-12-N1-GDO; PEG-23-N2-GDO; PEG-18-N3-GDO; PEG-23-N4-GDO; PEG-8-S1 -GDO; PEG-18-S2-GDO; PEG-12-S3-GDO; PEG-18-Ac1-GDO; PEG-12-Ac2-GDO; PEG-12-N1-GDM; PEG-12-N1-GDLO ;PEG-12-S3-GDM; PeG-12-S3-GDLO; PEG-12-Ac2-GDM; PEG-12-Ac2-GDLO; PEG-23-N1-GDL; PEG-12kN1-GDP; PEG-23 -Ac2-GDL and PEG-12-Ac2-GDP. GDLO means glycerol linolenate, and GDP means glycerol dipalmitate ester. Each of the compounds is a fluid at 25 ° C and has a stacking parameter Pa in the range of 0.830 to 0.869 and a Pv in the range of 0.872 to 0.924.

本發明進一步包括滿足其中所列舉之物理要求之已知脂質，其(例如)在25℃下為液體，且在20℃及37℃二者下自成型，且具有功能等效或等效堆疊參數，且當與水溶液組合時，在極少能量輸入或無能量輸入之情況下形成熱力學穩定之脂質體。 The present invention further encompasses known lipids that meet the physical requirements recited therein, which are, for example, liquid at 25 ° C and self-forming at both 20 ° C and 37 ° C with functional equivalent or equivalent stacking parameters And when combined with an aqueous solution, thermodynamically stable liposomes are formed with little or no energy input.

可用於本發明調配物之抗VEGF抗體包括任一已知抗VEGF抗體。該等抗體包括整個抗體或抗體片段，限制條件係其具有必需之抗VEGF生物學性質。在較佳實施例中，該抗體係具有必需之抗VEGF生物學及藥理學性質之抗體片段。美國專利第6,884,879號揭示可用於本發明之抗VEGF抗體。該等抗體包括具有包括以下之性質之人類化抗VEGF抗體及抗VEGF抗體變體：對VEGF之強結合親和力；抑制內皮細胞之VEGF促進之增生之能力及抑制VEGF誘導之血管生成之能力。較佳結合親和力(Kd)係不超過約5×10^-9M，且具有不超過約5nM之活體外抑制內皮細胞之VEGF誘導之增生之ED50值。該等抗體包括在A673活體內腫瘤模型中以5mg/kg之抗體劑量抑制至少約50%腫瘤生長之彼等。最佳抗體係以商標名稱LUCENTIS®(蘭尼單抗)出售，且經批准以玻璃體內調配物形式用於治療年齡相關黃斑退化及各種形式之黃斑水腫。術語「抗VEGF抗體」包括整個抗體以及抗體片段。可利用抗VEGF抗體治療之疾病範圍包括與血管生成或病理血管生成病狀相關之彼等疾病或病狀。該等包括癌症以及眼內新生血管症候群，例如增生性視網膜病變或年齡相關黃斑退化(AMD)、類風濕性關節炎及牛皮癬。儘管較佳投與途徑為對眼睛之局部治療，且較佳疾病模態為糖尿病黃斑水腫，但本文中所列舉之調配物亦可用於其他遞送模式(即，可注射；靜脈內輸注)且用於治療多種VEGF相關之疾病及病狀。 Anti-VEGF antibodies useful in the formulations of the invention include any of the known anti-VEGF antibodies. Such antibodies include whole antibodies or antibody fragments, with the proviso that they possess the requisite anti-VEGF biological properties. In a preferred embodiment, the anti-system has the necessary antibody fragments against the biological and pharmacological properties of VEGF. U.S. Patent No. 6,884,879 discloses an anti-VEGF antibody useful in the present invention. Such antibodies include humanized anti-VEGF antibodies and anti-VEGF antibody variants having the following properties: strong binding affinity for VEGF; ability to inhibit VEGF-promoted proliferation of endothelial cells and ability to inhibit VEGF-induced angiogenesis. Preferably, the binding affinity (Kd) is no more than about 5 x 10 ^-9 M and has an ED50 value that inhibits VEGF-induced proliferation of endothelial cells in vitro by no more than about 5 nM. Such antibodies include those that inhibit at least about 50% of tumor growth at a dose of 5 mg/kg of antibody in an in vivo tumor model of A673. The best anti-system is sold under the trade name LUCENTIS® (Lanibizumab) and is approved for use in the treatment of age-related macular degeneration and various forms of macular edema in the form of intravitreal formulations. The term "anti-VEGF antibody" includes whole antibodies as well as antibody fragments. The range of diseases that can be treated with anti-VEGF antibodies includes those diseases or conditions associated with angiogenesis or pathological angiogenic conditions. These include cancer and intraocular neovascular syndrome such as proliferative retinopathy or age-related macular degeneration (AMD), rheumatoid arthritis and psoriasis. Although the preferred route of administration is topical treatment of the eye, and the preferred disease mode is diabetic macular edema, the formulations listed herein can also be used in other modes of delivery (ie, injectable; intravenous infusion) and For the treatment of a variety of VEGF-related diseases and conditions.

抗VEGF抗體包括自編碼親代抗VEGF抗體之人類化變體之分離核酸產生之彼等，該親代抗體包含非人類可變結構域，其中該人類化變體結合人類VEGF，並具有6,884,879專利中所闡述並主張之彼等重鏈互補決定區胺基酸序列，該專利以引用的方式併入本文中。該抗VEGF抗體包括可使用具有編碼該等CDR胺基酸序列之核酸之載體及含有該等載體之分離宿主細胞產生的彼等。可培養宿主細胞以產生該等序列，且可自該等宿主細胞培養物回收人類化抗VEGF抗體。上文所列舉之分離核酸可進一步編碼具有含'879專利中所列舉之彼等序列之輕鏈互補決定區(CDR)之人類化變體。該人類化變體可包含具有顯示為'879專利中之SEQ ID NO：7之序列之重鏈可變結構域及具有顯示為SEQ ID NO：8之序列之輕鏈可變結構域。該人類化變體亦可包含'879專利中所顯示之SEQ ID NO：116之重鏈可變結構域序列及SEQ ID NO：115之輕鏈可變結構域序列。 Anti-VEGF antibodies include those produced from isolated nucleic acids encoding humanized variants of a parental anti-VEGF antibody comprising a non-human variable domain, wherein the humanized variant binds to human VEGF and has a patent of 6,884,879 The heavy chain complementarity determining region amino acid sequences set forth and claimed herein are incorporated herein by reference. The anti-VEGF antibody comprises those which can be produced using vectors having nucleic acids encoding the CDR amino acid sequences and isolated host cells containing such vectors. Host cells can be cultured to produce such sequences, and humanized anti-VEGF antibodies can be recovered from such host cell cultures. The isolated nucleic acids recited above may further encode humanized variants having light chain complementarity determining regions (CDRs) comprising the sequences set forth in the '879 patent. The humanized variant may comprise a heavy chain variable domain having the sequence set forth in SEQ ID NO: 7 of the '879 patent and a light chain variable domain having the sequence shown as SEQ ID NO: 8. The humanized variant may also comprise the heavy chain variable domain sequence of SEQ ID NO: 116 and the light chain variable domain sequence of SEQ ID NO: 115 as shown in the '879 patent.

美國專利第7,060,269號亦以引用的方式併入本文中。此專利主張並揭示蘭尼單抗。'269專利之技術方案1主張抑制受試者之VEGF誘導之血管生成之方法，該方法包含向該受試者投與有效量之人類化抗VEGF抗體，該人類化抗VEGF抗體以不超過約1×10^-8之Kd值結合人類VEGF，該人類化抗VEGF抗體包含SEQ ID NO：116之重鏈可變結構域序列及SEQ ID NO：115之輕鏈可變結構域序列。蘭尼單抗係經設計用於眼內使用之重組人類化IgG1 κ同型單株抗體片段。此單株抗體結合人類血管內皮生長因子A(VEGF-A)至並抑制其。蘭尼單抗具有約48,000道耳頓之分子量，並於含有四環素之營養培養基中之大腸桿菌表現系統中產生。此產物係以商標名稱LUCENTIS®市面有售，並以於單次使用玻璃小瓶中之不含防腐劑之滅菌溶液形式供應，其可遞送0.05mL之具有10mM組胺酸HCl、10% α,α海藻糖脫水物、0.01%聚山梨醇酯20且在pH為5.5之10mg/mL蘭尼單抗水溶液。 U.S. Patent No. 7,060,269 is incorporated herein by reference. This patent asserts and discloses ranibizumab. Technical Solution 1 of the '269 patent claims a method of inhibiting VEGF-induced angiogenesis in a subject, the method comprising administering to the subject an effective amount of a humanized anti-VEGF antibody, the humanized anti-VEGF antibody being no more than about The Kd value of 1 x 10 ^-8 binds to human VEGF, which comprises the heavy chain variable domain sequence of SEQ ID NO: 116 and the light chain variable domain sequence of SEQ ID NO: 115. Ranibizumab is a recombinant humanized IgG1 κ homologous antibody fragment designed for intraocular use. This monoclonal antibody binds to and inhibits human vascular endothelial growth factor A (VEGF-A). Ranibizumab has a molecular weight of about 48,000 Daltons and is produced in an E. coli expression system in a nutrient medium containing tetracycline. This product is marketed under the trade name LUCENTIS® and is supplied as a preservative-free sterile solution in a single-use glass vial that delivers 0.05 mL of 10 mM histidine HCl, 10% alpha, alpha Trehalose dehydrate, 0.01% polysorbate 20 and 10 mg/mL ranibizumab aqueous solution at pH 5.5.

蘭尼單抗係闡述於1999年發行之科學期刊中，於Journal of Molecular Biology(JMB)中由Chen等人以「Selection and Analysis of an Optimized Anti-VEGF Antibody：Crystal Structure of an Affinity-matured Fab in complex with Antigen」⁵(Chen等人，JMB,293：865-881(1999)為標題闡述。蘭尼單抗之重鏈及輕鏈序列在此文章中係命名為YO317，且係顯示於其中之圖1中。除此闡述以外，該文章亦提供關於此抗體片段與VEGF之結合親和力之數據(其中第870頁之表6)。已知蘭尼單抗結合至VEGF-A並抑制VEGF-A之生物活性，VEGF-A在眼部血管生成模型中引起新血管生成及洩漏。蘭尼單抗結合至VEGF-A並抑制其，並預防VEGF-A與內皮細胞表面上之VEGF受體相互作用，且從而減少新血管形成(血管生成)；血管洩漏及內皮細胞增生。醫藥上有效量之蘭尼單抗之投與抑制VEGF誘導之血管生成。術語抗VEGF抗體涵蓋全長抗體及抗體片段，例如Fab、Fab’、F(ab)₂及F_v，限制條件係該等片段藉由結合至人類VEGF顯示期望藥理學活性。蘭尼單抗與VEGF具有不超過約1×10^-8M、即約1.4×10^-10之結合親和力(Kd)(參見Chen等人，第870頁)。'269專利之圖10A及10B提供蘭尼單抗之輕鏈可變及重鏈可變結構域之序列(Chen等人中所顯示之Fab Y0317)。該等序列與'269專利中之SEQ ID NO：115及SEQ ID NO：116相同。 Lanibizumab is described in the 1999 issue of the scientific journal, in the Journal of Molecular Biology (JMB) by Chen et al. "Selection and Analysis of an Optimized Anti-VEGF Antibody: Crystal Structure of an Affinity-matured Fab in Complex with Antigen" ⁵ (Chen et al., JMB, 293:865-881 (1999) for the title. The heavy and light chain sequences of ranibizumab are named YO317 in this article and are shown in In Figure 1. In addition to this, the article also provides data on the binding affinity of this antibody fragment to VEGF (Table 6 on page 870). It is known that ranibizumab binds to VEGF-A and inhibits VEGF-A. Biological activity, VEGF-A causes neovascularization and leakage in ocular angiogenesis models. Ranibizumab binds to VEGF-A and inhibits it, and prevents VEGF-A from interacting with VEGF receptors on the surface of endothelial cells And thereby reducing neovascularization (angiogenesis); vascular leakage and endothelial cell proliferation. Administration of a pharmaceutically effective amount of ranibizumab inhibits VEGF-induced angiogenesis. The term anti-VEGF antibody encompasses full length antibodies and antibody fragments, eg Fab Fab ', F (ab) ₂ and F _v, based constraints such fragments. Ranibizumab to VEGF have no more than about 1 × 10 ^-8 M to human VEGF bound by displaying the desired pharmacological activity, i.e., about 1.4 Binding affinity (Kd) of ×10 ^-10 (see Chen et al., p. 870). Figures 10A and 10B of the '269 patent provide sequences of light chain variable and heavy chain variable domains of ranibizumab (Chen Fab Y0317) shown in et al. These sequences are identical to SEQ ID NO: 115 and SEQ ID NO: 116 of the '269 patent.

除以上文章及專利中所闡述之蘭尼單抗及其他抗VEGF抑制劑或藥物以外，其他抗VEGF或抗血管生成藥物亦可用於本發明調配物。儘管本發明者已發現為抗體片段之抗VEGF抗體在用於疾病及病狀之治療之脂質體調配物中較佳，該治療涉及向具有健康角膜或角膜之微量新血管生成但具有一些其他眼部病狀(例如，年齡相關黃斑退化或糖尿病黃斑水腫)之患者之眼睛局部施加，但包含整個抗VEGF抗體及自成型之熱力學穩定之脂質體之調配物亦可用於治療角膜新血管生成 及該等其他眼部疾病(在具有兩種疾病之患者中)，限制條件係角膜新血管生成容許或有助於較大整個抗體及其調配物進入。完整或整個抗VEGF抗體之實例係由Genentech以商標名稱AVASTIN®(貝伐珠單抗)出售。此抗體係抑制VEGF之生物活性之重組人類化IgG1抗體。其含有人類框架區及結合至VEGF之鼠類抗體之互補決定，且具有149kD之近似分子量。此抗體係於含有慶大黴素(Gentamycin)之營養培養基中之哺乳動物細胞(中國倉鼠卵巢)表現系統中產生。美國專利第6,054,297號(以引用的方式全文併入本文中)主張並揭示貝伐珠單抗或製造貝伐珠單抗之製程(參見技術方案1、6、7、8、9、10、12、29及30)。 In addition to the ranibizumab and other anti-VEGF inhibitors or drugs described in the above articles and patents, other anti-VEGF or anti-angiogenic drugs can also be used in the formulations of the present invention. Although the inventors have found that anti-VEGF antibodies which are antibody fragments are preferred in liposome formulations for the treatment of diseases and conditions, the treatment involves microangiogenesis with a healthy cornea or cornea but with some other eyes. Topical application to the eye of a patient with a condition (eg, age-related macular degeneration or diabetic macular edema), but a formulation containing the entire anti-VEGF antibody and self-forming thermodynamically stable liposomes can also be used to treat corneal neovascularization And such other ocular diseases (in patients with both diseases), the limiting condition is that corneal neovascularization allows or contributes to the entry of larger whole antibodies and their formulations. Examples of intact or whole anti-VEGF antibodies are sold by Genentech under the trade name AVASTIN® (bevacizumab). This anti-system inhibits the biological activity of VEGF by recombinant humanized IgG1 antibody. It contains the complementarity of the human framework region and the murine antibodies that bind to VEGF and has an approximate molecular weight of 149 kD. This anti-system is produced in a mammalian cell (Chinese hamster ovary) expression system in a nutrient medium containing gentamicin. U.S. Patent No. 6,054,297, the disclosure of which is hereby incorporated by reference in its entirety in its entirety, the entire disclosure of the disclosure of the disclosure of the disclosure of the disclosure of the disclosure of bevacizumab or the manufacture of bevacizumab (see technical solutions 1, 6, 7, 8, 9, 10, 12) , 29 and 30).

如經批准產物包裝插頁中所闡述，貝伐珠單抗係在活體外及活體內分析系統中結合至人類血管內皮生長因子並抑制其生物活性之重組人類化單株IgG1抗體。此抗體含有結合至VEGF之鼠類抗體之人類框架區⁶(參見L.G.Presta等人，(1997)Cancer Res.57：4593-99)。貝伐珠單抗之分子量係約149千道耳頓。此紙張揭示人類化F(ab)抗體片段之可變結構域「F(ab)1-12」之相互作用。貝伐珠單抗具有衍生自鼠類抗體之序列之非人類CDR以及可變結構域中於輕鏈(V_L)之46位及重鏈(V_H)之49位、69位、71位、73位、78位及94位處之框架取代，該等框架取代與F(ab)-12之相應位置處所顯示之取代相同，如Presta等人之圖1中所顯示。Presta等人具有關於貝伐珠單抗之分子特徵及結合特徵之資訊。 Bevacizumab is a recombinant humanized monoclonal IgG1 antibody that binds to human vascular endothelial growth factor and inhibits its biological activity in an in vitro and in vivo assay system as set forth in the approved product packaging insert. This antibody contains human framework region ⁶ of murine antibodies that bind to VEGF (see LG Presta et al. (1997) Cancer Res. 57:4593-99). The molecular weight of bevacizumab is about 149 thousand Daltons. This paper reveals the interaction of the variable domain "F(ab)1-12" of the humanized F(ab) antibody fragment. Bevacizumab has 46 derived from non-human heavy chain variable domains and CDR sequences of the murine antibody light chain (V _L) of (V _H) of 49, 69, 71, Substituted by the 73, 78 and 94 frames, these frame substitutions are identical to those shown at the corresponding positions of F(ab)-12, as shown in Figure 1 of Presta et al. Presta et al. have information on the molecular characteristics and binding characteristics of bevacizumab.

如先前所陳述，除蘭尼單抗及貝伐珠單抗以外，其他已知抗VEGF抗體或抗血管生成藥物亦可用於本發明。本發明之抗VEGF抗體係如上文關於其所引用之專利參考文獻中所闡述製備。通常，將編碼該抗體之分離核酸；包含該核酸之載體可操作連接至由經該載體轉化之宿主細胞識別之對照序列；具有該載體之宿主細胞全部共同用於在 培養該等細胞並收集並純化該抗體後產生所關注抗體之製程。可將任一適宜醫藥賦形劑添加至該抗體中，且亦可根據需要使該抗體凍乾。「抗VEGF抗體」包括各種形式，且可為具有完整人類Fc區之全長抗體或抗體片段(例如，Fab、Fab’或F(ab’)₂)。適於與本文中所揭示之脂質組合形成脂質體調配物之其他抗血管生成藥物包括哌加他尼(pegaptanib)或依那西普(etanercept)(TNF抑制劑)。在後一情形中，此調配物可用於治療各種自體免疫疾病或病狀。依那西普係以商標名稱Enbrel®出售，其係用於治療類風濕性、幼年型類風濕性及牛皮癬關節炎、斑塊狀牛皮癬及關節黏連性脊髓炎。其他適宜藥物包括舒尼替尼，即VEGF及PDGF受體蛋白激酶及血管生成抑制劑(以名稱舒癌特(SUTENT®)出售之2-羥吲哚)，且其係闡述並主張於美國專利第6,573,293號中(該專利案之內容已以引用的方式併入本文中)；或FOVISTA^TM(以前稱為E10030)，即血小板源生長因子B(PDGF-B)之調節劑(1.5mg/.5mg蘭尼單抗)。組合使用之其他適宜藥物包括干擾素-α-2a或替西羅莫司(Temsirolimus)或其他mTOR抑制劑(例如)雷帕黴素(rapamycin)。可組合使用之用於眼部疾病之藥物類別亦包括蛋白酶體抑制劑、自體吞噬抑制劑、類視色素、溶酶體抑制劑、熱休克反應激活劑、Hsp90伴隨蛋白抑制劑、蛋白運輸抑制劑、糖苷酶抑制劑、酪胺酸激酶抑制劑及組蛋白去乙醯酶抑制劑。該等藥物可單獨用於脂質體調配物，或可以與抗VEGF化合物或抗體之組合成調配物形式使用，或可以依序組合形式且較佳以局部調配物形式使用。當組合FOVISTA^TM(0.03-3.0mg/眼睛，與0.5mg蘭尼單抗或其他抗VEGF化合物組合)(及/或具有PDGF抑制活性之其他藥物)與蘭尼單抗時，較佳適應症係治療年齡相關黃斑退化。阿柏西普(Aflibercept)(2.0mg/0.05mL)(Eylea^TM)亦可單獨或與蘭尼單抗或其他活性成份組合用於脂質體調配物。本發明進一步包括本文中所列舉包含FOVISTA及阿 柏西普及/或任一其他抗血管生成藥物之局部脂質體調配物，限制條件係至少一種活性成份係與本發明之熱力學穩定之自成型脂質體摻和/組合。FOVISTA(針對PDGF-B之適配體)亦已知且在美國專利公開案第2012/0100136號中闡述為「拮抗劑A」，該專利公開案之內容已以引用的方式全文併入本文中。拮抗劑A之合成係闡述於'136公開案中之實例4中(亦參見其中之圖7)。當所闡述之個別VEGF拮抗劑及PDGF拮抗劑中至少一種藥劑與本文中所闡述形成熱力學穩定之自成型脂質體之脂質調配在一起時，其中每一者亦包括於本發明之範圍內。具有本文中所列舉之活性成份中之任一者或任一組合之局部調配物優於藉由玻璃體內注射投與之藥物或其組合。另外，向眼睛局部施加用於眼部疾病相對於全身性經口投與較佳。可用於本發明之組合物包括(以脂質體調配物或脂質體調配物及/或任一組合形式之其他調配物之形式)(a)(PDGF抑制劑)ARC-127、拮抗劑A、拮抗劑B、拮抗劑C、拮抗劑D、1B3抗體、CDP860、IMC-3G3、伊馬替尼(imatinib)、162.62抗體、163.31抗體、169.14抗體、169.31抗體、αR1抗體、2A1E2抗體、M4TS.11抗體、M4TS.22抗體、A10、佈雷菲德菌素A(brefeldinA)、舒尼替尼、Hyb 120.1.2.1.2抗體、Hyb 121.6.1.1.1抗體、Hyb 127.5.7.3.1抗體、Hyb 127.8.2.2.2抗體、Hyb 1.6.1抗體、Hyb 1.11.1抗體、Hyb 1.17.1抗體、Hyb 1.18.1抗體、Hyb 1.19.1抗體、Hyb 1.23.1抗體、Hyb 1.24抗體、Hyb 1.25抗體、Hyb 1.29抗體、Hyb 1.33抗體、Hyb 1.38抗體、Hyb 1.39抗體、Hyb 1.40抗體、Hyb 1.45抗體、Hyb 1.46抗體、Hyb 1.48抗體、Hyb 1.49抗體、Hyb 1.51抗體、Hyb 6.4.1抗體、F3抗體、人類化F3抗體、C1抗體、人類化C1抗體、6.4抗體、抗mPGDF-C山羊IgG抗體、C3.1抗體、5-甲基-7-二乙基胺基-s-***并(1,5-a)嘧啶、干擾素、魚精蛋白、PDGFR-B1單株抗體、PDGFR-B2單株抗體、6D11單株抗體、Sis單株抗體、PR7212單株抗體、PR292單 株抗體、HYB9610單株抗體、HYB 9611單株抗體、HYB 9612單株抗體、HYB 9613單株抗體、苯磺酸4-(2-(N-(2-甲醯胺基-吲哚)胺基乙基)酯、4-(2-(N-(-2-甲醯胺吲哚)胺基乙基)-磺醯基脲、CGP 53716、人類抗體g162、吡唑并[3,4-g]喹喏啉、6-[2-(甲基胺甲醯基)苯基硫烷基]-3-E-[2-(吡啶-2-基)乙烯基]-吲唑、1-{2-[5-(2-甲氧基-乙氧基)-苯并咪唑-1-基]-喹啉-8-基}-六氫吡啶-4-基胺、4-{4-[N-(4-硝基苯基)胺甲醯基]-1-六氫吡嗪基]-6,7-二甲氧基喹唑啉、4-胺基-5-氟-3-(6-(4-甲基-六氫吡嗪-1-基)-1H-苯并咪唑-2-基)1H-喹啉-2-酮、(4-第三丁基苯基){4-[(6,7-二甲氧基-4-喹啉基)氧基]苯基}甲酮、5-甲基-N-[4-(三氟甲基)苯基]-4-異噁唑甲醯胺、反式-4-[(6,7-二甲氧基喹喏啉-2-基)胺基]環己醇、(Z)-3-[(2,4-二甲基-5-(2-側氧基-1,2-二氫吲哚-3-亞基甲基)-1H-吡咯-3-基)-丙酸、5-(5-氟-2-側氧基-1,2-二氫吲哚-3-亞基甲基)-2,4-二甲基-1H-吡咯-3-甲酸、1-(4-氯苯胺基)-4-(4-吡啶基甲基)酞嗪、N-{4-(3-胺基-1H-吲唑-4-基)苯基-N”-(2-氟-5-甲基苯基)脲、1,2-二甲基-7-(2-噻吩)咪唑并[5,4-g]喹喏啉、1,2-二甲基-6-苯基-咪唑并[5,4-g]喹喏啉、1,2-二甲基-6-(2-噻吩)咪唑并[5,4-g]喹喏啉、AG1295、AG1296、3-芳基喹啉、4-吡啶基-2-芳基嘧啶、索拉菲尼(sorafenib)、MLN518、PKC412、AMN107、蘇拉明(suramin)、新黴素或其醫藥上可接受之鹽；及(b)(VEGF抑制劑)蘭尼單抗、貝伐珠單抗、阿柏西普、KH902VEGF受體-Fe融合蛋白、2C3抗體、ORA102、哌加他尼、貝伐西尼(bevasiranib)、鈍端貝伐西尼、SIRNA-027、紫花前胡素(decursin)、紫花前胡醇(decursinol)、鬼臼苦素(picropodophyllin)、香膠樹脂酮(guggulsterone)、PLG101、類花生酸LXA4、PTK787、帕唑帕尼(Pazopanib)、阿西替尼(axitinib)、CDDO-Me、CDDO-Imm、紫草素、β羥基異戊醯基紫草素、神經節苷脂GM3、DC101抗體、Mab 25抗體、Mab73抗體、4A5抗體、4E10抗體、5F12抗體、VA01抗體、BL2 抗體、BECG相關蛋白、sFLT01、sFLT02、肽B3、TG100801、索拉菲尼、G6-31抗體或抑制VEGF相關之血管生成之其他化合物。除抗體以外，適宜於治療眼部疾病或病狀(包括眼睛前部之疾病，例如角膜疾病)或因手術切口、創傷或潰瘍而必需之癒合的其他基於蛋白質之活性成份包括(例如)人類生長激素或其他已知激素肽或其變體。 As previously stated, in addition to ranibizumab and bevacizumab, other known anti-VEGF antibodies or anti-angiogenic drugs are also useful in the present invention. The anti-VEGF anti-systems of the invention are prepared as set forth above in the patent references cited therein. Typically, an isolated nucleic acid encoding the antibody; a vector comprising the nucleic acid operably linked to a control sequence recognized by a host cell transformed with the vector; a host cell having the vector is used in common for culturing the cells and collecting The process of purifying the antibody produces the antibody of interest. Any suitable pharmaceutical excipient can be added to the antibody, and the antibody can also be lyophilized as needed. An "anti-VEGF antibody" includes various forms and can be a full length antibody or antibody fragment (eg, Fab, Fab' or F(ab') ₂ ) having an intact human Fc region. Other anti-angiogenic drugs suitable for combination with the lipids disclosed herein to form liposome formulations include pegaptanib or etanercept (TNF inhibitor). In the latter case, the formulation can be used to treat a variety of autoimmune diseases or conditions. Enesip is sold under the trade name Enbrel® for the treatment of rheumatoid, juvenile rheumatoid and psoriatic arthritis, plaque psoriasis and joint adhesion myelitis. Other suitable drugs include sunitinib, a VEGF and PDGF receptor protein kinase and an angiogenesis inhibitor (2-hydroxyindole sold under the name SUTENT®), which is described and claimed in the US patent. No. 6,573,293 (the contents of which are incorporated herein by reference); or FOVISTA ^(TM) (formerly known as E10030), a modulator of platelet-derived growth factor B (PDGF-B) (1.5 mg/. 5mg ranibizumab). Other suitable drugs for use in combination include interferon-α-2a or temsirolimus or other mTOR inhibitors such as rapamycin. The drug classes that can be used in combination for ocular diseases also include proteasome inhibitors, autophagy inhibitors, retinoids, lysosomal inhibitors, heat shock response activators, Hsp90 concomitant protein inhibitors, and protein transport inhibition. Agents, glycosidase inhibitors, tyrosine kinase inhibitors and histone deacetylase inhibitors. The drugs may be used alone in liposome formulations, or may be used in combination with an anti-VEGF compound or antibody, or may be used in sequential combination and preferably in the form of a topical formulation. When the combination FOVISTA ^TM (0.03-3.0mg / eye, in combination with 0.5mg ranibizumab or other anti-VEGF compounds) (and / or with other pharmaceutically active inhibiting the PDGF) and ranibizumab, preferred indications based Treatment of age-related macular degeneration. Aflibercept (Aflibercept) (2.0mg / 0.05mL) (Eylea TM) may also be used alone or in combination ranibizumab or other active ingredients for the liposome formulations. The invention further encompasses a topical liposome formulation comprising FOVISTA and Abcy's universal/or any other anti-angiogenic drug as set forth herein, the limitation being at least one active ingredient and the thermodynamically stable self-forming liposomes of the invention Blending/combination. FOVISTA (aptamer for PDGF-B) is also known and described in the US Patent Publication No. 2012/0100136 as "Antagonist A", the contents of which are hereby incorporated by reference in its entirety herein. . The synthesis of antagonist A is illustrated in Example 4 of the '136 publication (see also Figure 7 therein). When at least one of the individual VEGF antagonists and PDGF antagonists described is formulated with the lipids forming the thermodynamically stable self-forming liposomes as described herein, each of which is also included within the scope of the invention. Topical formulations having any one or any combination of the active ingredients recited herein are preferred over drugs or combinations thereof administered by intravitreal injection. In addition, local application to the eye for ocular disease is preferred over systemic oral administration. Compositions useful in the present invention include (in the form of liposome formulations or liposome formulations and/or other formulations in any combination) (a) (PDGF inhibitor) ARC-127, antagonist A, antagonism Agent B, antagonist C, antagonist D, 1B3 antibody, CDP860, IMC-3G3, imatinib (imatinib), 162.22 antibody, 163.31 antibody, 169.14 antibody, 169.31 antibody, αR1 antibody, 2A1E2 antibody, M4TS.11 antibody, M4TS.22 antibody, A10, brefeldin A, sunitinib, Hyb 120.1.2.1.2 antibody, Hyb 121.6.1.1.1 antibody, Hyb 127.5.7.3.1 antibody, Hyb 127.8.2.2 .2 Antibody, Hyb 1.6.1 Antibody, Hyb 1.11.1 Antibody, Hyb 1.17.1 Antibody, Hyb 1.18.1 Antibody, Hyb 1.19.1 Antibody, Hyb 1.23.1 Antibody, Hyb 1.24 Antibody, Hyb 1.25 Antibody, Hyb 1.29 Antibody, Hyb 1.33 Antibody, Hyb 1.38 Antibody, Hyb 1.39 Antibody, Hyb 1.40 Antibody, Hyb 1.45 Antibody, Hyb 1.46 Antibody, Hyb 1.48 Antibody, Hyb 1.49 Antibody, Hyb 1.51 Antibody, Hyb 6.4.1 Antibody, F3 Antibody, Humanized F3 Antibody, C1 antibody, humanized C1 antibody, 6.4 antibody, anti-mPGDF-C goat IgG antibody, C3.1 antibody, 5-methyl-7- Ethylamino-s-triazolo (1,5-a) pyrimidine, interferon, protamine, PDGFR-B1 monoclonal antibody, PDGFR-B2 monoclonal antibody, 6D11 monoclonal antibody, Sis monoclonal antibody, PR7212 monoclonal antibody, PR292 monoclonal antibody, HYB9610 monoclonal antibody, HYB 9611 monoclonal antibody, HYB 9612 monoclonal antibody, HYB 9613 monoclonal antibody, 4-(2-(N-(2-carbamide)) Base-oxime aminoethyl ester, 4-(2-(N-(2-carbamimidoxime)aminoethyl)-sulfonyl urea, CGP 53716, human antibody g162, pyrazole And [3,4-g]quinoxaline, 6-[2-(methylamine-mercapto)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]- Carbazole, 1-{2-[5-(2-methoxy-ethoxy)-benzimidazol-1-yl]-quinolin-8-yl}-hexahydropyridin-4-ylamine, 4 -{4-[N-(4-nitrophenyl)amine-carbamoyl]-1-hexahydropyrazinyl]-6,7-dimethoxyquinazoline, 4-amino-5-fluoro -3-(6-(4-methyl-hexahydropyrazin-1-yl)-1H-benzoimidazol-2-yl)1H-quinolin-2-one, (4-t-butylphenyl) {4-[(6,7-Dimethoxy-4-quinolinyl)oxy]phenyl}methanone, 5-methyl-N-[4-(trifluoromethyl)phenyl]- 4-isoxazolecarbamamine, trans-4-[(6,7-dimethoxyquinoxalin-2-yl)amino] ring Alcohol, (Z)-3-[(2,4-dimethyl-5-(2-o-oxy-1,2-dihydroindol-3-ylidenemethyl)-1H-pyrrole-3- Propionate, 5-(5-fluoro-2-oxo-1,2-dihydroindole-3-ylidenemethyl)-2,4-dimethyl-1H-pyrrole-3- Formic acid, 1-(4-chloroanilino)-4-(4-pyridylmethyl)pyridazine, N-{4-(3-amino-1H-indazol-4-yl)phenyl-N" -(2-fluoro-5-methylphenyl)urea, 1,2-dimethyl-7-(2-thiophene)imidazo[5,4-g]quinoxaline, 1,2-dimethyl -6-phenyl-imidazo[5,4-g]quinoxaline, 1,2-dimethyl-6-(2-thiophene)imidazo[5,4-g]quinoxaline, AG1295, AG1296 , 3-arylquinoline, 4-pyridyl-2-arylpyrimidine, sorafenib, MLN518, PKC412, AMN107, suramin, neomycin or pharmaceutically acceptable Salt; and (b) (VEGF inhibitor) ranibizumab, bevacizumab, aboxicept, KH902VEGF receptor-Fe fusion protein, 2C3 antibody, ORA102, pegaptanib, bevacizine ( Bevasiranib), blunt-end bevacizini, SIRNA-027, decursin, decursinol, picropodophyllin, guggulsterone, PLG101, peanut Acid LXA4 PTK787, Pazopanib, axitinib, CDDO-Me, CDDO-Imm, shikonin, beta hydroxyisoamyl shikonin, ganglioside GM3, DC101 antibody, Mab 25 antibody, Mab73 antibody, 4A5 antibody, 4E10 antibody, 5F12 antibody, VA01 antibody, BL2 antibody, BECG-related protein, sFLT01, sFLT02, peptide B3, TG100801, sorafenib, G6-31 antibody or VEGF-related angiogenesis Other compounds. In addition to antibodies, other protein-based active ingredients suitable for the treatment of ocular diseases or conditions, including diseases in the anterior eye, such as corneal diseases, or necessary healing due to surgical incisions, wounds or ulcers include, for example, human growth. Hormone or other known hormone peptides or variants thereof.

脂質體調配物係藉由以任一順序遵循以下一般步驟來製備：(1)提供含有抗VEGF抗體及/或上文所闡述之其他活性物之水溶液；(2)將能夠形成脂質體之熱力學穩定之自成型脂質添加至步驟(1)之該水溶液中；及(3)視情況添加醫藥上可接受之賦形劑。可利用製備脂質體懸浮液調配物之製程之任一變化形式，包括組合抗VEGF抗體(或VEGF抑制劑或PDGF抑制劑)與脂質且然後添加水溶液或將每一成份分開添加至水溶液中。基於預期遞送途徑(例如，局部等)製備該懸浮液，且亦基於該途徑選擇其他賦形劑。載劑、穩定劑及/或賦形劑包括緩衝液，例如磷酸鹽、檸檬酸鹽或其他無機酸；抗氧化劑，例如抗壞血酸及/或甲硫胺酸；防腐劑；低分子量多肽；蛋白質，例如明膠、血清白蛋白或免疫球蛋白；親水聚合物，例如PVP；胺基酸；單糖或二糖或其他碳水化合物；螯合劑；糖；鹽形成抗衡離子；非離子型表面活性劑及諸如此類。該脂質體調配物亦可呈溶液之形式。 Liposomal formulations are prepared by following the general procedure in either order: (1) providing an aqueous solution containing an anti-VEGF antibody and/or other actives as set forth above; (2) thermodynamics capable of forming liposomes A stable self-forming lipid is added to the aqueous solution of step (1); and (3) a pharmaceutically acceptable excipient is added as appropriate. Any variation of the process for preparing a liposome suspension formulation can be utilized, including combining an anti-VEGF antibody (or VEGF inhibitor or PDGF inhibitor) with a lipid and then adding an aqueous solution or separately adding each component to the aqueous solution. The suspension is prepared based on the intended route of delivery (eg, topical, etc.) and other excipients are also selected based on this route. Carriers, stabilizers and/or excipients include buffers such as phosphates, citrates or other inorganic acids; antioxidants such as ascorbic acid and/or methionine; preservatives; low molecular weight polypeptides; Gelatin, serum albumin or immunoglobulin; hydrophilic polymers such as PVP; amino acids; monosaccharides or disaccharides or other carbohydrates; chelating agents; sugars; salts forming counterions; nonionic surfactants and the like. The liposome formulation can also be in the form of a solution.

該等調配物可用於治療VEGF相關之疾病及病症。欲利用本文所闡述之調配物治療之較佳疾病或病狀係眼部疾病。如上文所闡述，本發明之較佳疾病或病狀係治療糖尿病黃斑水腫。如上文所參考，DME係由一系列生物化學及細胞事件造成，該等事件最終引起漸進洩漏及滲出，從而導致視網膜增厚及黃斑中央之一個視神經盤直徑內形成硬滲出物。雷射光凝係主要治療，且可有效以約505預防中度視覺喪失之風險[ETDRSRG,1985]。雷射光凝改良閱讀線得分，但具有相關併發症，例如瘢痕漸進增大、中央盲點、對比敏感度降低及色覺 受損。 Such formulations are useful for treating diseases and conditions associated with VEGF. The preferred disease or condition to be treated with the formulations described herein is an ocular condition. As indicated above, the preferred disease or condition of the invention is the treatment of diabetic macular edema. As referred to above, DME is caused by a series of biochemical and cellular events that ultimately cause progressive leakage and exudation, resulting in thickening of the retina and formation of hard exudate within the diameter of one of the optic discs in the center of the macula. Laser photocoagulation is the primary treatment and can effectively prevent the risk of moderate visual loss at approximately 505 [ETDRSRG, 1985]. Laser photocoagulation improves reading line scores, but has associated complications such as progressively enlarged scars, central blind spots, reduced contrast sensitivity, and color vision Damaged.

脂質體調配物可廣泛地治療與VEGF及/或PDGF相關之腫瘤或視網膜病症或任一其他眼部疾病或病狀，此取決於特定活性成份。抗VEGF抗體抑制一或多種由VEGF引起之生物學活性。治療應用涉及向需要特定疾病或病狀治療之患者投與之醫藥上可接受之劑型。適宜劑型儘管較佳為局部，但亦可包括藉由呈濃注形式之靜脈內方式或藉由連續輸注；肌內、腹膜內、腦脊髓內、皮下、關節內、滑膜內、鞘內、經口或藉由吸入之投與。另外，該等抗體調配物亦可藉由腫瘤內、瘤周、病灶內或灶周途徑來投與。適於利用抗體調配物治療之腫瘤性疾病包括各種癌，包括乳癌、肺癌、胃癌、食管癌、結腸直腸癌、肝癌、卵巢癌、含睪丸細胞之卵巢腺瘤、子宮頸癌、子宮內膜癌、子宮內膜增生、子宮內膜組織異位症、纖維肉瘤、絨毛膜癌、頭頸癌、鼻咽癌、喉癌、肝胚細胞癌、卡波西氏肉瘤(Kaposi’s sarcoma)、黑色素瘤、皮膚癌、血管瘤、海綿狀血管瘤、血管母細胞瘤、胰臟癌及其他類型之癌症。VEGF相關之非腫瘤性病狀包括類風濕性關節炎、牛皮癬、動脈粥樣硬化、糖尿病及其他增生性視網膜病變(包括早產兒視網膜病變)、晶狀體後纖維組織增生、新生血管性青光眼、年齡相關黃斑退化、糖尿病黃斑水腫及其他形式之黃斑水腫、甲狀腺增生(包括格雷氏病(Graves’ disease))、角膜及其他組織移植、慢性發炎、肺發炎、腎炎症候群、子癇前症、腹水、心包積液及肋膜積液。利用較佳局部調配物治療之較佳病狀或疾病係糖尿病黃斑水腫。所投與之劑量及投與頻率將取決於疾病之類型及嚴重性及特定患者之狀態。例如，抗VEGF抗體可以1μg/kg至約50mg/kg或約0.1mg/kg至20mg/kg之劑量範圍向需要治療之患者投與。用於治療DME且關於蘭尼單抗之局部調配物之較佳劑量方案係闡述於本文實例3中。活性成份之濃度及量可視特定患者及治療天數而變，且每天或週 或月所提供之量亦可視患者之反應以及視覺敏銳度及視網膜增厚之改良之體徵而變。 Liposomal formulations can broadly treat tumor or retinal disorders associated with VEGF and/or PDGF or any other ocular disease or condition, depending on the particular active ingredient. Anti-VEGF antibodies inhibit one or more of the biological activities caused by VEGF. Therapeutic applications involve the administration of a pharmaceutically acceptable dosage form to a patient in need of treatment for a particular disease or condition. Suitable dosage forms, although preferably local, may also be included by intravenous means in the form of a bolus or by continuous infusion; intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, By mouth or by inhalation. In addition, such antibody formulations may also be administered by intratumoral, peritumoral, intralesional or periventricular routes. Tumor diseases suitable for treatment with antibody formulations include various cancers including breast cancer, lung cancer, gastric cancer, esophageal cancer, colorectal cancer, liver cancer, ovarian cancer, ovarian adenoma containing sputum cells, cervical cancer, endometrial cancer , endometrial hyperplasia, endometrial tissue dysplasia, fibrosarcoma, choriocarcinoma, head and neck cancer, nasopharyngeal carcinoma, laryngeal cancer, hepatic blast cell carcinoma, Kaposi's sarcoma, melanoma, skin Cancer, hemangioma, cavernous hemangioma, hemangioblastoma, pancreatic cancer, and other types of cancer. VEGF-related non-neoplastic conditions include rheumatoid arthritis, psoriasis, atherosclerosis, diabetes and other proliferative retinopathy (including retinopathy of prematurity), post-lens fibrous tissue hyperplasia, neovascular glaucoma, age-related macular Degenerative, diabetic macular edema and other forms of macular edema, thyroid hyperplasia (including Graves' disease), corneal and other tissue transplantation, chronic inflammation, lung inflammation, renal inflammation, pre-eclampsia, ascites, pericardial effusion And pleural effusion. The preferred condition or disease treated with a preferred topical formulation is diabetic macular edema. The dose and frequency of administration will depend on the type and severity of the disease and the condition of the particular patient. For example, the anti-VEGF antibody can be administered to a patient in need of treatment at a dose ranging from 1 [mu]g/kg to about 50 mg/kg or from about 0.1 mg/kg to 20 mg/kg. Preferred dosage regimens for the treatment of DME and for topical formulations of ranibizumab are set forth in Example 3 herein. The concentration and amount of active ingredient may vary depending on the particular patient and the number of treatment days, and daily or weekly The amount provided by the month may also vary depending on the patient's response and the signs of improvement in visual acuity and retinal thickening.

出於治療DME或其他VEGF相關之眼部病狀之目的之理想治療模態可係導致迅速且持久之視覺改良者。目前用於治療DME之其他治療形態包括選擇性PKCβ抑制劑(魯伯斯塔(ruboxistaurin))；類固醇(曲安奈德(triamcinolone acetonide)、丙酮氟洛皮質醇(fluocinolone acetonide))；VEGF抑制劑(貝伐珠單抗；蘭尼單抗及哌加他尼-可注射劑)及玻璃體切除術。本發明脂質體調配物提供較(例如)目前市售玻璃體內調配物顯著改良之局部治療方案。本發明調配物可與其他已知治療組合用於本文中所列舉及/或上文所闡述之眼部或VEGF相關之疾病或病狀，且限制條件係不存在任何禁忌。該等治療方案或治療方法包括(例如)siRNA分子(例如貝伐西尼)以及包括本發明所利用之熱力學穩定之自成型脂質體之適當遞送媒劑。 The ideal therapeutic modality for the purpose of treating DME or other VEGF-related ocular conditions can result in rapid and long-lasting visual improvement. Other therapeutic modalities currently used to treat DME include selective PKC beta inhibitors (ruboxistaurin); steroids (triamcinolone acetonide, fluocinolone acetonide); VEGF inhibitors ( Bevacizumab; ranibizumab and pegaptanib-injectables) and vitrectomy. The liposome formulations of the present invention provide a topical treatment regimen that is significantly improved over, for example, current commercial intravitreal formulations. The formulations of the invention may be used in combination with other known therapies for the diseases or conditions associated with the eye or VEGF recited herein and/or as set forth above, and the limitations are not contraindicated. Such therapeutic regimens or methods of treatment include, for example, siRNA molecules (e.g., bevacziron) and suitable delivery vehicles including thermodynamically stable self-forming liposomes utilized in the present invention.

可單獨或與任一其他活性成份組合用於脂質體調配物之眼用類固醇包括***、膚輕鬆(fluocinolone)、氯替潑諾(loteprednol)、二氟潑尼酯(difluprednate)、氟米龍(fluorometholone)、去氫皮質醇(prednisolone)、甲羥松(medrysone)、曲安奈德、利美索龍(rimexolone)及其各種鹽形式。其他眼用抗炎劑(例如NSAID)亦可用於脂質體調配物。視活性成份而定，可利用除熱力學穩定之自成型脂質體以外或作為其之替代方案之其他脂質體。 Ophthalmic steroids which may be used alone or in combination with any other active ingredient for liposome formulations include dexamethasone, fluocinolone, loteprednol, difluprednate, flumi Fluorometholone, prednisolone, medrysone, triamcinolone acetonide, rimexolone and various salt forms thereof. Other ophthalmic anti-inflammatory agents (eg, NSAIDs) can also be used in liposome formulations. Depending on the active ingredient, other liposomes other than or as an alternative to thermodynamically stable self-forming liposomes may be utilized.

以下實例意欲進一步闡釋本發明之某些實施例，且並非限制性： The following examples are intended to further illustrate certain embodiments of the invention, and are not limiting:

實例Instance 實例1-A Example 1-A 脂質體及蘭尼單抗之溶液Liposomes and solutions of ranibizumab

獲得含有0.5mg濃度為10mg/mL之蘭尼單抗(0.05mL)之小瓶。將0.015克PEG-12甘油二肉豆蔻酸酯(PEG-12 GDM)Qsomes^TM添加至 此溶液中(PEG後面之數字指示PEG鏈中之C₂H₄O亞單位之數量)。使用1.45mL由磷酸鹽、氯化鈉及硬脂酸聚羥氧40酯組成之緩衝溶液將此脂質體懸浮液之體積稀釋至1.5mL之最終體積，以提供於脂質體懸浮液中0.333mg/mL之蘭尼單抗濃度及約1%(10mg/mL)之脂質百分比。添加過硼酸鈉(0.28mg/mL)作為防腐劑。1mL之此懸浮液相當於20滴。每一滴含有大約17μg蘭尼單抗。 A vial containing 0.5 mg of ranibizumab (0.05 mL) at a concentration of 10 mg/mL was obtained. The 0.015 g PEG-12 glyceryl dimyristate (PEG-12 GDM) Qsomes ^TM this solution was added (hereinafter the PEG chain PEG numbers indicate the number of C ₂ H ₄ O subunits of). The volume of the liposome suspension was diluted to a final volume of 1.5 mL using 1.45 mL of a buffer solution consisting of phosphate, sodium chloride and polyoxyl 40 stearate to provide 0.333 mg/ml of liposome suspension. The concentration of ranibizumab in mL and the percentage of lipid in about 1% (10 mg/mL). Sodium perborate (0.28 mg/mL) was added as a preservative. 1 mL of this suspension is equivalent to 20 drops. Each drop contained approximately 17 μg of ranibizumab.

藉由組合15mL硬脂酸聚羥氧40酯、氯化鈉、磷酸二氫鈉及磷酸氫二鈉溶液與5mL過硼酸鈉溶液來製備該緩衝溶液(V=20mL,pH 5.5)。然後如上文所直接闡述利用1.45mL之此溶液。眼用脂質體懸浮液調配物中之每一賦形劑之濃度為0.142mg/mL(磷酸氫二鈉)；6.7mg/mL(磷酸二氫鈉)；50mg/mL(硬脂酸聚羥氧40酯)；5.1mg/mL(氯化鈉)；0.333mg/mL(蘭尼單抗)；10mg/mL(PEG-12 GDM)及2.8mg/mL(過硼酸鈉)。可利用HCl或NaOH調整pH，且亦可利用低分子量胺基酸或有機酸。圖3顯示至少兩種類型之脂質體(Qusomes®)，該等脂質體係在將脂質與水溶液混合時形成(來自Biozone Laboratories網站)。 The buffer solution (V = 20 mL, pH 5.5) was prepared by combining 15 mL of polyoxyl 40 stearate, sodium chloride, sodium dihydrogen phosphate and disodium hydrogen phosphate solution with 5 mL of sodium perborate solution. Then 1.45 mL of this solution was used as explained directly above. The concentration of each excipient in the ophthalmic liposome suspension formulation was 0.142 mg/mL (disodium hydrogen phosphate); 6.7 mg/mL (sodium dihydrogen phosphate); 50 mg/mL (polyoxyl stearate) 40 ester); 5.1 mg/mL (sodium chloride); 0.333 mg/mL (ranibizumab); 10 mg/mL (PEG-12 GDM) and 2.8 mg/mL (sodium perborate). The pH can be adjusted using HCl or NaOH, and a low molecular weight amino acid or an organic acid can also be utilized. Figure 3 shows at least two types of liposomes (Qusomes®) formed upon mixing lipids with an aqueous solution (from the Biozone Laboratories website).

實例2-Example 2 兔角膜中之擴散室研究 Study on diffusion chamber in rabbit cornea

使用下文所闡述之方法生成施加至兔角膜之脂質體調配物之擴散室數據。概言之，在10分鐘、20分鐘及30分鐘及1小時、2小時、3小時、4小時、5小時、6小時及24小時處獲取樣品。該等數據顯示局部施加之脂質體蘭尼單抗調配物在34℃下進入兔角膜之眼前房液中之顯著滲透速率。在脂質體調配物中，在3小時處開始鑑別蘭尼單抗，且其在投與後保持存在長達24小時，與先前針對非脂質體調配物在兔中所報導之7天及14天相對(數據未顯示，參見Chen等人，Eye London 2011 Nov；25(11)：1504-11)。在具有水平流之玻璃Valia-Chen室中實施研究。水以34℃之溫度再循環。將膜置於室之接面之間，且在此實例 中，使用兔角膜作為該膜。用3.2mL之鹽水溶液填充受體室，以模擬眼睛之前面部分中之眼前房液之內容物。供體室係提供有3mL包含蘭尼單抗及熱力學穩定之自成型脂質之眼用調配物。不斷攪動擴散室。在各種時間點處自受體室收集樣品-獲取400μL樣品並每次用400μL之鹽水溶液替代。在以下時間點處獲取樣品：10min；20min；30min；1hr；2hr；3hr；4hr；5hr；6hr及24hr。早在第3個小時時藉由HPLC檢測蘭尼單抗。使用Lucentis®作為HPLC標準之對照溶液。亦使用電泳評價脂質體調配物穿過兔角膜膜，且結果與HPLC數據一致。 Diffusion chamber data for liposome formulations applied to rabbit corneas were generated using the methods set forth below. In summary, samples were taken at 10 minutes, 20 minutes, and 30 minutes and at 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, and 24 hours. These data show the significant rate of permeation of the locally applied liposomal ranibizumab formulation into the anterior chamber fluid of the rabbit cornea at 34 °C. In liposome formulations, ranibizumab was identified at 3 hours and it remained present for up to 24 hours after administration, compared to 7 days and 14 days previously reported for rabbits in non-liposomal formulations. Relative (data not shown, see Chen et al., Eye London 2011 Nov; 25(11): 1504-11). The study was carried out in a glass Valia-Chen chamber with horizontal flow. The water is recirculated at a temperature of 34 °C. Place the membrane between the junctions of the chamber, and in this example In the middle, a rabbit cornea is used as the film. The receptor chamber was filled with 3.2 mL of saline solution to simulate the contents of the anterior chamber fluid in the anterior portion of the eye. The donor compartment is provided with 3 mL of an ophthalmic formulation containing ranibizumab and thermodynamically stable self-forming lipids. Stir the diffusion chamber constantly. Samples were collected from the receptor chamber at various time points - 400 μL samples were taken and replaced with 400 μL of saline solution each time. Samples were taken at the following time points: 10 min; 20 min; 30 min; 1 hr; 2 hr; 3 hr; 4 hr; 5 hr; 6 hr and 24 hr. Ranibizumab was detected by HPLC as early as the third hour. Lucentis® was used as a control solution for the HPLC standard. Liposomal formulations were also assessed by electrophoresis through rabbit corneal membranes and the results were consistent with HPLC data.

實例3-Example 3 對患有DME之患者之先導性臨床研究Pilot clinical study of patients with DME

每天利用每3小時1滴之調配物將患有DME之患者治療6次/天(6×/天)，並持續2週。蘭尼單抗之總劑量/天係6×17ug或102ug。在此2週時期後整個6週內見到平均中央窩厚度(CFT)之損失之改良及視覺敏銳度之增加(參見圖1及圖2)。在第8週，發生視網膜厚度之增加及視覺敏銳度之降低，且在第10週，以2滴/天(34ug/天)再起始治療。在第14週，觀測到OCT及BCVA改良之清晰趨勢(參見圖1及2)。亦使用相同方案治療2個其他患者。所有三個患者之結果係呈現於圖4至7中，並顯示CFT及VA相對於對照之改良。 Patients with DME were treated 6 times per day (6 x/day) with 1 drop of formulation every 3 hours for 2 weeks. The total dose of ranibizumab / day is 6 x 17ug or 102ug. Improvements in the loss of mean central fossa thickness (CFT) and an increase in visual acuity were seen throughout the 6 weeks after this 2-week period (see Figures 1 and 2). At week 8, an increase in retinal thickness and a decrease in visual acuity occurred, and at week 10, treatment was initiated at 2 drops/day (34 ug/day). At week 14, a clear trend in OCT and BCVA improvement was observed (see Figures 1 and 2). The same regimen was also used to treat 2 other patients. The results for all three patients are presented in Figures 4 to 7 and show improvements in CFT and VA relative to controls.

實例4-Example 4 使用曲安奈德對患有DME之患者進行之先導性臨床研究Pilot clinical study of patients with DME using triamcinolone acetonide

在單一中心、開放標籤之先導性研究中，患有DME之合格患者接受包含去炎松(triamcinolone，TA)之局部調配物。3位患有涉及黃斑中央之DME之患者(平均年齡58歲，在53歲至64歲範圍內)之總共3隻眼睛在研究眼睛之最佳矯正視力(BCVA)使用ETDRS測試65個至40個字母。在十二(12)週之控制治療時期期間在患者醒著時，引導其每2小時將含有133ug(微克)之TA之1滴施加於研究眼睛中(6次)。主要結果量度包括3個月時之主要端點，例如眼部及全身性不良事件之頻率 及嚴重性以及中央窩厚度(CFT)相對於基線之變化，如藉由光學同調斷層掃描術(OCT)隨時間來量測。次要結果係相對於基線BCVA得分隨時間之變化、根據眼底照相(FP)之ETDRS視網膜病變嚴重性相對於基線具有>3步進展之患者之比例、根據螢光血管攝影(FA)解決洩漏之患者之比例及黃斑雷射治療隨時間之需求。以與蘭尼單抗調配物類似之方式自市售起始材料製備該TA調配物。 In a single center, open label pilot study, eligible patients with DME received a topical formulation containing triamcinolone (TA). A total of 3 eyes of 3 patients with DME in the center of the macula (mean age 58 years, ranging from 53 to 64 years old) were tested for best corrected visual acuity (BCVA) using the ETDRS test 65 to 40 letter. During the twelve (12) weeks of controlled treatment period, one ounce of 133 ug (micrograms) of TA was administered to the study eyes (6 times) every 2 hours while the patient was awake. The primary outcome measure included the primary endpoint at 3 months, such as the frequency of ocular and systemic adverse events. And severity and changes in foveal thickness (CFT) relative to baseline, as measured by optical coherence tomography (OCT) over time. Secondary outcomes were based on changes in baseline BCVA score over time, proportion of patients with >3 progression of baseline ETDRS retinopathy based on fundus photography (FP), and resolution of leakage based on fluorescence angiography (FA) The proportion of patients and the need for macular laser treatment over time. The TA formulation was prepared from commercially available starting materials in a manner similar to the ranibizumab formulation.

去炎松+1%脂質體眼用懸浮液 De-inflammatory pine + 1% liposome ophthalmic suspension

最終調配物係滅菌水性懸浮液。其含量係如以下： The final formulation is a sterile aqueous suspension. Its content is as follows:

PEG-12-GDM：脂質體；二醯基甘油-聚乙二醇(PEG 12)、甘油二肉豆蔻酸酯(GDM)。 PEG-12-GDM: liposome; dimercaptoglycerol-polyethylene glycol (PEG 12), glyceryl dimyristate (GDM).

* TA係最終產物。其係微粉化TA(大約12mm)且不含防腐劑。 * TA is the final product. It is micronized TA (about 12 mm) and contains no preservatives.

去炎松+1%脂質體眼用懸浮液之製備方案 Preparation scheme of triamcinolone acetonide liposome ophthalmic suspension

1. 將蒸餾水之最終體積之40%置於燒杯中，且將其加熱至70℃ 至80℃。 1. Place 40% of the final volume of distilled water in a beaker and heat it to 70 °C To 80 ° C.

2. 添加羥丙基甲基纖維素，並停止混合，直至達到室溫且其變成澄清且均質之混合物為止。 2. Add hydroxypropyl methylcellulose and stop mixing until room temperature is reached and it becomes a clear and homogeneous mixture.

3. 對其進行高壓處理，且在滅菌後使其達到室溫，同時攪拌。 3. Treat it with high pressure and allow it to reach room temperature while sterilizing while stirring.

4. 將蒸餾水之最終體積之40%置於另一燒杯中。添加並混合，直至逐個完全溶解以下試劑為止：a)磷酸二氫鈉b)磷酸氫二鈉c)EDTA d)氯化鈉e)聚山梨醇酯80 4. Place 40% of the final volume of distilled water in another beaker. Add and mix until the following reagents are completely dissolved: a) sodium dihydrogen phosphate b) disodium hydrogen phosphate c) EDTA d) sodium chloride e) polysorbate 80

5. 在10%之剩餘體積之水中，以50%添加苯紮氯銨並混合，直至完全納入為止。在溶解後，將此新溶液添加至含有磷酸鹽、EDTA、氯化鈉及聚山梨醇酯80之上文溶液中。為滅菌，藉由0.22μm膜過濾。 5. Add 10% benzalkonium chloride in 50% of the remaining volume of water and mix until completely incorporated. After dissolution, this new solution was added to the above solution containing phosphate, EDTA, sodium chloride, and polysorbate 80. For sterilization, it was filtered through a 0.22 μm membrane.

6. 混合羥丙基甲基纖維素之滅菌溶液與含有該等鹽及防腐劑苯紮氯銨之另一滅菌溶液，並混合直至得到澄清均質混合物為止。 6. Mix the sterilized solution of hydroxypropyl methylcellulose with another sterile solution containing the salt and the preservative benzalkonium chloride, and mix until a clear homogeneous mixture is obtained.

7. 將乙酸去炎松添加至具有緩衝液及苯紮氯銨之溶液中，並攪拌直至完全納入為止。 7. Add triamcinolone acetate to the solution with buffer and benzalkonium chloride and stir until fully incorporated.

8.將脂質體添加至此混合物中，並在15分鐘期間利用磁性攪拌器攪拌，以獲得最終懸浮液。 8. Liposomes were added to this mixture and stirred with a magnetic stirrer during 15 minutes to obtain the final suspension.

9. 將該懸浮液封裝於專用眼睛滴管中。每一滴管瓶含有1.5mL之此去炎松眼用懸浮液。 9. Enclose the suspension in a dedicated eye dropper. Each dropper bottle contains 1.5 mL of this triamcinolone eye suspension.

結果： 在脂質體調配物中包含TA之局部調配物在患有涉及中央之臨床顯著之DME之患者中的使用具有良好耐受性。未報導眼部或全身性不良事件。在第2個月，所有三個患者之CFT相對於基線而降 低。三個患者中之兩者具有至少100um之CFT降低。在第3個月，所有三個患者顯示視覺敏銳度改良。患者中之一者得到>15個字母。 Results: Topical formulations comprising TA in liposome formulations are well tolerated in use in patients with centrally significant clinically significant DME. No eye or systemic adverse events were reported. At 2 months, the CFT of all three patients decreased relative to baseline. Two of the three patients had a CFT reduction of at least 100 um. In the third month, all three patients showed visual acuity improvement. One of the patients received >15 letters.

儘管已詳細並參照其具體實施例闡述所主張發明，但熟習此項技術者將顯而易見，可對所主張發明進行各種變化及修改，而不背離其精神及範圍。因此，例如，熟習此項技術者僅使用常規實驗即可認識到或能夠確定所主張發明之許多可尚未明確闡述之實施例。該等實施例屬於本發明之範圍。 Although the invention has been described in detail and by reference to the specific embodiments thereof, it will be understood that Thus, for example, those skilled in the art will recognize, <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; These embodiments are within the scope of the invention.

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Claims (5)

一種局部眼用調配物之用途，其係用於製備治療患者眼後段疾病之藥物，其中該局部眼用調配物包含脂質體及類固醇，其中該脂質體係自成型之熱力學穩定之脂質體。 A use of a topical ophthalmic formulation for the manufacture of a medicament for treating a disease in the posterior segment of the eye, wherein the topical ophthalmic formulation comprises a liposome and a steroid, wherein the lipid system is self-forming thermodynamically stable liposomes. 如請求項1之用途，其中該眼後段疾病係糖尿病黃斑水腫。 The use of claim 1, wherein the disease in the posterior segment of the eye is diabetic macular edema. 如請求項1之用途，其中該類固醇係選自曲安奈德。 The use of claim 1, wherein the steroid is selected from the group consisting of triamcinolone acetonide. 如請求項3之用途，其中曲安奈德係以50ug/每滴調配物至150ug/每滴之劑量範圍投與，並每2小時向眼睛局部施加1滴，4次/天至6次/天。 The use of claim 3, wherein the triamcinolone acetonide is administered in a dose range of 50 ug per drop to 150 ug per drop, and 1 drop is applied to the eye every 2 hours, 4 times/day to 6 times/day. . 如請求項1之用途，其中該患者顯示改善之中央窩厚度及視覺敏銳度。 The use of claim 1, wherein the patient exhibits improved central fossa thickness and visual acuity.